CHAPTER 1

INTRODUCTION
THE TRACER CONCEPT and helium ions and free electrons. With time
A brief historical view the temperature decreased and after 700 million
years the electrons were able to attach to the
Big Bang ions forming neutral atoms and the plasma was
transformed to a large cloud of hydrogen and
helium gas. In the neutral gas gravity was able
to locally condense the gas, a process that again
increased the temperature, and the first stars, the
red giants were born. The temperature and the
density in the stars increased the probability of
other nuclear reactions like the formation of
beryllium-8 by two helium nuclides. This
nucleus could then pick up another helium ion
(or α-particle) to form carbon-12. In steps of
four mass units (2 protons and 2 neutrons) light
elements were formed, oxygen-16, neon-20,
manganese-24, sulphur-32 etc in a process
called fusion. Other reactions such as neutron
capture, splitting of the target nucleus in nuclear
reactions and radioactive decay contributed to
create other combinations of proton and
neutrons.
All matter in the universe has its origin in an
event called the “Big Bang”, a cosmic explosion The fusion process worked for elements up to
releasing an enormous amount of energy about iron but for more heavy elements other
14 billion years ago. Particles like protons and mechanisms took over. The iron nucleus picked
neutrons, the building stones of the nucleus, up neutrons creating instable proton and neutron
were condensed as free particles during the first combinations that decayed emitting a negative
seconds. However, it took several minutes until -
the decreasing temperature of the expanding charge in the form of a beta particle (β ). A new
universe reached a level that allowed the element (cobalt) with an extra proton in the
formation of particle combinations like nucleus was then created. The cobalt nucleus
deuterium (heavy hydrogen) as well as helium captured new neutrons and formed nickel nuclei
(see figure below). via radioactive decay. In such a way, step by
32
step, all the heavy elements we know were
16 S
15 Heavy ion fusion in red super giants
31
P created.
28 29 30
14 Si Si Si
12C-fusion 27
13 Al
When the red giants grow old they exploded.
n
O-fusio

24 25 26
12 n Mg Mg Mg
Number of protons

11
23
Na All their content of heavy material was spread
10 α 20Ne 21 Ne 22Ne around in universe. By gravity other stars
16

19
9 F
8 He-fusion in
16
O O O n
17 18 picked it up and formed planets. Actually, all
7 red giants
14 15
N N matter around us, including our-selves, was
α C C
12 13 n
6
10 11
once created in one of these red stars.
5 B B
4 α 8 Be 9 Be
3
4
6 7
Li Li Billion years have passed since our planet
2 He
Tellus was formed. Most of the unstable proton-
n p+n d (deuterium), d+d He
1 p d
0 n
H-fusion in gas clouds and stars neutron combinations once created has
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 undergone transformations (radioactive decay)
Number of neutrons into stable combinations. However, some with
very long half-lives remain: potassium-40, lead-
For several 100 million years the universe was a 204, thorium-232 and the natural occurring
large plasma composed by hydrogen, deuterium isotopes of uranium.

1
The pioneers
These “glowing” pieces after the cosmic
cookery event were discovered by Henri
Becquerel 1896 and were further investigated
by Marie Curie and her husband Pierre Curie. It
was the scientific sensation of that time, which
still has great implications in our lives.
Suddenly several new elements were discovered
that emitted radiation of several types. First was
polonium named after Poland, Marie Curie’s
country of birth. The second was radium, which
was soon found to have properties useful in
medicine (cancer treatment). However, some
elements were chemically identical with already
known elements, such as lead, but emitted this
strange new radiation. This was puzzling for the
scientists at that time. They had only vague
knowledge about the structure of the atom, and
the neutron was still to be discovered. Soddy, in
1913, introduced the concept of isotopes
(Greek: iso= same, tope= place). Isotopes
occupied the same place in the periodical
system, were chemically identical, but differed
from each other in atomic weight and emission
of radioactivity. When Chadwick in 1932
discovered the neutron this concept was given a
rational explanation.
Antoine Henri Becquerel Marie Curie
Frederick Soddy George de Hevesy
2
Almost at the same time (1913), George de
Hevesy demonstrated the practical implication
of the isotopic theory. He and his colleagues
used a radioactive isotope of lead, 210Pb, as a
tracer (or indicator) when they studied the
solubility of sparingly soluble lead salts. Hevesy
was also the first to apply the radioactive tracer
technique in biology when he investigated lead
uptake in plants (1923) using 212Pb. Only one
year later, Blumengarten and Weiss carried out
the first clinical study. They injected 212Bi into
one arm of a patient and measured the arrival
time in the other arm. From this study, they
concluded that the arrival time was prolonged in
patients with heart disease.
Induced radioactivity
So far, nature was the supplier of the radioactive
nuclides that were used. Isotopes of uranium
and thorium generated a variety of radioactive
heavy elements such as lead, but radioactive
isotopes of light elements were not known.
Marie Curie’s daughter Irène, together with her
husband Frédéric Joliot took the next step.
Alpha-emitting sources had for long been used
to bombard different elements. Rutherford
studied e.g. the deflection of alpha particles in
gold foils and discovered the small, charged
nucleus of the atom. But Joliot-Curie also
showed that the alpha particles induced
radioactivity in the bombarded foil (in their case
aluminum foil). The induced radioactivity had a
half-life of about 3 minutes.
Frédéric Joliot Irène Joliot-Curie
Correctly, they identified the radiation to be
emitted from 30P created in the nuclear reaction.
27
Al (α, n) 30P

They also concluded that “These elements and
similar ones may possibly be formed in
different nuclear reactions with other
bombarding particles: protons, deuterons and
neutrons. For example, 13N could perhaps be
formed by capture of a deuteron in 12C,
followed by the emission of a neutron.”
Ernest Orlando Lawrence Enrico Fermi
The same year this was proved to be true by
Ernest Lawrence in Berkeley, California and
Enrico Fermi, Rome. Lawrence had built a
cyclotron capable of accelerating deuterons up
to about 3 MeV and it was by pure chance that
this group had not yet discovered induced
radioactivity. He soon reported the production
of 13N with a half-life of 10 minutes. After this
the cyclotron was used to produce several other
biologically important radionuclides such as
11
C, 32P and 22Na.
Fermi realized that the neutron was
advantageous for radionuclide production. Since
it has no charge, it could easily enter into the
nucleus and induce a nuclear reaction. He
immediately made a strong neutron source by
sealing up 232Ra-gas with beryllium powder in a
glass vial. The alpha particle emitted from 232Ra
caused a nuclear reaction in beryllium and a
neutron was emitted.
9
Be (α, n) 12C
Fermi started a systematical search by
irradiating all available elements in the periodic
system with fast and slow neutrons to study the
creation of induced radioactivity. From
hydrogen to oxygen, no radioactivity was
observed in their targets, but in the ninth
element, fluorine, their hopes were fulfilled. In
3
the following weeks, they bombarded some 60
elements and found induced radioactivity in 40
of them. They also observed that the lighter
elements were usually transmuted into
radionuclides of a different chemical element,
whereas heavier elements appeared to yield
radioisotopes of the same element as the target.
These new discoveries excited the scientific
community. From having been a rather limited
technique, the radioactivity tracer principle
could suddenly be applied in a variety of fields
and especially in life sciences. De Hevesy
immediately started to study the uptake and
elimination of 32P-phosphate in various tissues
of rats and demonstrated, for the first time, the
kinetics of vital elements in living creatures. 128I
was applied in the diagnosis of thyroid disease.
This was the start of the radiotracer technology
in biology and medicine, as we know it today.
One early cyclotron-produced nuclide was 11C.
It is of special importance since carbon is
fundamental in life science. 11C has a half-life
of only 20 minutes but by setting up a chemical
laboratory close to the cyclotron, organic
compound labeled with 11C can be obtained in
large amounts. Photosynthesis was studied
using 11CO2 and the fixation of carbon
monoxide in humans by the use of inhalation of
11
CO. However, 20 minutes was short and the
use of 11C was limited to the most rapid
biochemical reactions. One must remember that
the radio-detectors used at that time were
primitive and that the chemical synthetic and
analytic tools were not adapted to such short
times. A search to find a more long-lived
isotope of carbon resulted 1939 in the discovery
of 14C produced in the nuclear reaction
13
C (d, p) 14C
However, 14C produced this way was of limited
use since the radionuclide could not be
separated from the target.
During the bombardments, a bottle of
ammonium nitrate solution had been standing
close to the target. By pure chance, it was
discovered that this bottle also contained 14C,
which had been produced in the reaction
14
N (n, p) 14C
 may say that reactor-produced radionuclides are
The deuterons used in the bombardment consist
most suitable for laboratory work, whereas
of one proton and one neutron with a binding
accelerator-produced radionuclides are more
energy of about 2 MeV. When high-energy
clinically useful. Some of the most used
deuterons hit a target, it is likely that the
radionuclides in nuclear medicine, such as 111In
binding between the particles breaks and that a
and 201Tl, are accelerator produced. When using
free neutron is created in what is called a
very short-lived radionuclides (11C, 13N, 15O and
“stripping reaction”. The bottle with ammonium 18
F) for positron emission tomography (PET)
nitrate had thus unintentionally been neutron
the need for an in-house accelerator is obvious.
irradiated. Since no carbon was present in the
bottle (except small amounts from solved air-
Radioactive nuclides with short half-life may
borne carbon dioxide) the 14C produced this
play an important role in the future laboratory
way was of high specific radioactivity. It was
use of the tracer technique. We know that
also very easy to separate from the target. In the
biological systems, such as pheromones, may
nuclear reaction, a “hot” carbon atom was
work at extremely low concentrations. The
created, which formed 14CO2 in the solution. By
radiotracer technique, as we know it today, is
simply bubbling air through the bottle, the 14C
often not useful to study such systems since it is
was released from the target.
not sensitive enough. The drawback with 14C is
its very long half-life of 5370 years. If we have
The same year, and in the same place, tritium
one million 14C-labeled molecules, in the mean
was discovered by deuteron irradiation of water.
only one will decay in an experiment lasting a
One of the pioneers Martin Kamen stated:
few days. Usually, the practical detection limits
“Within a few months, after the scientific world
are in the pico-mole area. When using the more
had somewhat ruefully concluded that
short-lived 11C, almost all labeled molecules
development of tracer techniques would be
contribute to the signal during an experiment
seriously handicapped because useful
lasting one hour. The sensitivity increases
radioactive tracers for carbon, hydrogen, oxygen
considerably to the femto-mole or even atto-
and nitrogen did not exist, 14C and 3H were
mole level.
discovered, and the situation greatly improved”.
This is indeed true. The role of 3H- and 14C-
DEFINITION OF IONIZING RADIATION
labeled tracers in biochemical research is a story
in itself.
Ionizing radiation is high-energy particles or
photons that can cause ionization, in other
Before the Second World War, the cyclotron
words, cause the emission of electrons from
was the main producer of radionuclides since
atoms or molecules. Subatomic particles with a
the neutron sources at that time were very weak.
mass (such as electrons, protons and neutrons)
But with the development of the nuclear reactor,
transfer their kinetic energy, in different
the situation changed. Suddenly, a tremendously
processes, to the orbit electrons of the atom.
strong neutron source was available, which
Photons (electromagnetic radiation) with
easily could produce almost unlimited amounts
wavelength of atomic dimensions or less behave
of radioactive nuclides including such
as discrete ”packets” of energy (quanta). These
biological important elements as 3H, 14C, 32P,
35 quanta, in different processes, also transfer their
S and clinically interesting radionuclides as
60 energy to matter creating ionization.
Co (for external radiotherapy) and 131I for
nuclear medicine. After the war, a new industry
In 1895, Wilhelm Conrad Roentgen discovered
was born which could deliver a variety of
X-rays, the first man made ionization radiation.
radiolabeled compounds at a reasonable price.
When fast electrons are slowed down in dense
material, they create radiation of electro-
However, accelerator-produced nuclides have a
magnetic nature similar to that of ordinary
special character, which makes them differ from
visible light. However, these quanta, roentgen
reactor-produced nuclides. Today, their
or X-rays, have considerably higher energy than
popularity is increasing again. Generally, one

4
light. This type of continuous radiation is also
called bremsstrahlung.
The direction of the
field is perpendicular
to the plane of the
page. The heavy,
positive α −particle is
deflected slightly to
the left whereas the
light negative β-
particle is deflected
markedly to the right.
Figure 1 A collimated The uncharged γ -
photons being
radium source in a
magnetic field. insensitive to the
magnetic field carry
straight on.
In 1896, Henri Becquerel demonstrated that
uranium mineral emits penetrating radiation
with properties similar to X-rays. The couple,
Marie and Pierre Curie, showed in 1898 that
certain uranium minerals contained a previously
unknown element, radium, which emits intense
ionizing radiation of different types. A magnetic
field can separate the radium radiation into three
components (Figure 1).
On closer analysis, it became apparent that
α−radiation was identical with helium nuclei,
β-radiation consisted of electrons, and
γ-radiation was of electromagnetic nature.
Shortly afterwards, ionizing radiation was used
in medicine for diagnostic radiology and the
irradiation of tumors. However, at that time
there were little knowledge about the effects of
radiation on matter, how radiation transferred
energy to the region irradiated and how to
quantify radiation. Ionizing radiation is of
course invisible and humans have no sensory
organ that can detect it. Consequently, a series
of tragic events occurred in the early decades of
the use of ionization radiation. The need for
measuring techniques (dosimetry) and
protective procedures for those working with
ionizing radiation was soon perceived.
5
Subsequent development was rapid. The
technology to accelerate charged particles to
high energy was developed. In the 1930’s, a
new ionizing particle, the neutron, was
discovered. Like roentgen and gamma radiation,
it had no charge. It was soon demonstrated that
the neutron caused nuclear reactions including
nuclear fission in heavy elements.
During the1940’s, physicists learnt to control
the fission process, which in nuclear weapons
can liberate instantaneously enormous amounts
of energy in the form of ionizing radiation. In
nuclear reactors, nuclear fission can be
controlled and used to produce energy and large
amounts of radioactive material. The explosion
of nuclear weapons, especially in the biosphere,
disperses enormous amounts of radioactive
material, which resulted in many people being
exposed to radiation. Nuclear power plants also
contributed to an increase in radioactive
nuclides in the environment even if during
normal operation the increase is small.
HISTORICAL SUMMARY
1895 Discovery of X-rays
1896 Discovery of natural radioactivity
1900 - Period of serious personal injuries
1920 amongst radiologists due to lack of
knowledge.
1920 National and international radiation
protection standards begin to be
instituted.
1930 The neutron and induced radioactivity
discovered. Construction of particle
accelerators begins.
1939 Nuclear fission discovered
1945 Nuclear bombs used (Hiroshima and
Nagasaki).
1950- Development of nuclear reactors
1960- Use of reactors for production of
energy, development of trace element
techniques, increased use of radiation in
medicine, research and industry
Energy of ionizing radiation
When ionizing radiation (in the form of high-
energy photons, neutrons or charged particles)
collides with matter, the full energy or part of it
can be transferred to the matter in various types
of interaction. It is by such energy transfer that
radiation exerts its damaging effect on living
creatures. Part of the transmitted energy causes
ionization or excitation in the material (Figure
2). Ionization means that electrons in the matter
receive so much additional energy that they are
liberated completely from the atom or molecule
to which they are bound. By excitation, the
additional energy is insufficient for the electrons
to be liberated from the molecule.
Figure 2 Ionizing
radiation transfer
energy to the atom's
orbital electrons. They
can either be displaced
to a distal orbit
(excitation) or they can
be free electrons
(ionization).
The electron is displaced only to an orbit with a
lower binding energy. The atom or molecule
does not change its charge number but the
process may change its chemical properties.
+1 volt
e-, electron
Ee= 1 eV
ve= 600 km/s
e+, proton
Ep=1 eV
vp=14 km/s
0
Figure 3 An electron is accelerated from the
negative towards the positive electrode. At
arrival, the electron has acquired a kinetic
energy of 1 eV. A particle with positive charge,
e.g. a proton, undergoes a similar event from
the positive to the negative electrode. However,
since the proton mass is about 2000 times the
mass of the electron, the proton velocity is
considerably lower.
6
To cause ionization or excitation, it is essential
that the incoming radiation has certain energy.
This is expressed as electron volt eV. One eV is
the kinetic energy an electron or a proton
receives after acceleration in a potential drop of
1 volt (Figure 3).
Generally, charged and non-charged particles
can be allotted a certain kinetic energy
expressed in eV or
keV (kiloelectron volt = 103 eV),
MeV (megaelectron volt = 106 eV) and
GeV (gigaelectron volt = 109 eV).
X-ray and gamma radiation does not have mass
or kinetic energy in the real sense. However,
when their wavelength diminishes and begins to
approximate the dimensions of the atom and the
nucleus, the electromagnetic radiation can be
regarded as particles (photons or radiation
quanta). Each photon has certain energy (E)
E=hν
where ν is the frequency of the electromagnetic
oscillations of the radiation, h is "Planck's
constant", and E is energy in eV.
On average, about 30 eV are needed to cause
one ionization in most materials. We define
ionizing radiation to have energy higher than
100 eV. Radiation with lower energy is called
non-ionizing radiation.
PRODUCTION OF IONIZING
RADIATION
In electromagnetic fields, charged particles can
be accelerated to such energies that they can
penetrate matter. Important types of accelerators
are:
a for acceleration of electrons
1 X-ray tubes (0 - 300 keV)
2 Linear accelerators (2 - 100 MeV)
3 Betatrons (15 - 100 MeV)
4 Microtrons (5 - 50 MeV)
b for acceleration of heavy, charged
particles (protons, heavier ions)
1 Van de Graaff accelerator (1-25 MeV)
 2 Linear accelerator (1 - 800 MeV)
3 Cyclotron (1 - 1,000 MeV)
4 Synchrotron (1 - 300 GeV)
Secondary ionizing radiation arises when the
accelerated particles meet a radiation target.
a X-ray photons, when for example
electrons are decelerated in an X-ray tube.
Radiation protection when working with X-
ray equipment is dealt with in Chapter 5.
b Protons, neutrons, photons or other
radiation particles when protons, heavier
particles or photons cause nuclear reactions.
and beam current (number of particles per unit
of time or in microamperes). From the radiation
protection point of view, an essential feature of
accelerators is that radiation production can be
turned off easily. The only risk involved is the
possible occurrence of induced radioactivity.
Radioactive sources, which are the commonest
source of radiation in the laboratory, do not
have this convenient attribute.
Other examples are neutron and gamma
radiation, which arise as a result of nuclear
fission processes in a fission reactor.

Figure 4 Cyclotron that accelerates protons to

17 MeV. Generally used for radionuclide
production.

Figure 5 Modern diagnostic X-ray laboratory

for tomographic information.

Important parameters specifying an accelerator's

performance are the type of accelerated particles
(electrons, protons etc.), maximum particle
energy (most often given in keV, MeV or GeV)

7
 CHAPTER 2
NUCLEAR STABILITY AND INSTABILITY
Of these expressions only the isotope concept is
ORGANIZATION OF MATTER generally used. It is important to understand that
whenever we use the expression isotope, we
Chemists learnt during the late 19th century to must always relate it to a specific element or a
organize chemical knowledge into the periodic group of elements, e.g. isotopes of carbon (e.g.
11
system. Radioactivity, when it was discovered, C, 12C, 13C and 14C) or isotopes of the
conflicted with that system. Suddenly various halogens.
samples, apparently with the same chemical
NUCLIDE CHART
behavior, were found to have different physical
qualities such as half-life, emitted type of protons = neutrons
100 On this side
radiation and energy. The concept of isotopes or
neutron deficient

Number of protons
elements occupying the same place in the 80 nuclides which
periodic system (from the Greek: iso- = same
decay by β+
and topos = place) was introduced by Soddy 60 and EC
1913, but a complete explanation had to wait
for the discovery of the neutron (Chadwick 40 On this side
1932). neutron excess
20 nuclides which
The periodic system was organized according to decay by β-
the number of protons in the nucleus. This 0
related to the number of surrounding electrons 0 50 100 150
that determined the chemistry of the element. Number of neutrons
Since the number of neutrons in the nucleus is
of importance as well, the physicist’s way to Figure 1 Chart of nuclides. The black dots
organize matter, the nuclide chart, is somewhat represent 279 naturally existing combinations
different. Each proton-neutron combination is of protons and neutrons (stable or almost stable
identified in this chart. nuclides). Around this stable line are about
2300 proton/neutron-combinations that are
A proton-neutron combination is identified by radioactive.
A
ZX 9
17
F
18
F
19
F
20
F
21
F
22
F
23
F
24
F
25
F
13 14 15 16 17 18 19 20 21 22 23 24
8 O O O O O O O O O O O O
Number of protons

X = element name (e. g. C for carbon) 7

12
N 13
N 14
N 15
N 16
N 17
N 18
N 19
N 20
N 21
N 22
N 23
N
9 10 11 12 13 14 15 16 17 18 19 20
Z = number of protons in the nucleus 6
8
C C
10
C
11
C
12
C
13
C
14
C
15
C C
17
C C C
5 B B B B B B B B
N = number of neutrons in the nucleus 4
7 8
Be Be Be
9 10 11
Be Be Be
12 14
Be
A = mass number (A=Z+N) 3
6 7
Li Li Li Li
8 9 11
Li
3 4 6 8
2 He He He He
1 2 3
1 H H H
The expression above is over-determined. If the 0 n
element name X is known, so is the number of 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
protons in the nucleus, Z. Usually, the following Number of neutrons
simplified expression is used.
A
X Figure 2 A part of the nuclide chart where the
lightest elements are shown. The dark squares
Some relations of the numbers of protons and
represent stable nuclides that are important for
neutrons have special names such as
our existence. Nuclides to the left of the stable
Isotopes = number of proton is constant
ones are radionuclides deficient in neutrons and
(Z = constant)
those to the right, rich in neutrons.
Isotones = number of neutrons is constant
(N = constant)
The force that keeps the nucleons (protons and
Isobars = mass number is constant
neutrons) together in the nucleus is called the
(A = constant)
“strong force” or the “nuclear force”. Other

8
forces, e.g. the Coulomb force, tend to separate
the charged particles in the nucleus. In the
stable and in the radioactive nucleus, there is a
balance between binding and separating forces.
The interplay between the forces is illustrated in
Figure 4. A proton and a neutron are held
together by the strong force and create a stable
isotope of hydrogen, deuterium. Two protons do
not create a stable system since the Coulomb
force exceeds the nuclear force.
electron
proton
neutron
Figure 3 Here, three isotopes of the element
hydrogen are illustrated using a simple atomic
model. 1H has only one proton, deuterium 2H
has a proton and a neutron in its nucleus, and
tritium 3H has a proton and two neutrons.
Isotopes behave in the same way chemically
since it is the electron shell that determines
what happens in chemical reactions. Physically
they are completely different.
Between two neutrons, no Coulomb force is
present and one should expect this system to be
stable. However, this is not the case since
besides the strong force and the Coulomb force
a symmetry rule exist saying that a higher
nuclear stability is achieved when there are
about the same number of protons and neutrons
in the nucleus. This symmetry rule prevents two
neutrons from sticking together, but allows two
neutrons and a proton to form a radioactive
nucleus, 3H.
A useful measure is the binding energy that
keeps the nucleons together. If the energy of 2.2
MeV is added to a deuterium nucleus, the two
nucleons are parted as a free proton and a free
neutron. Another way to express this is to
measure the differences in weight. The
deuterium nucleus is somewhat lighter than the
sum weight of the free neutron and proton. This
mass difference can be converted into energy
using the Einstein formula E = mc2 and is found
to be 2.2 MeV.
9
proton neutron
Figure 4 Binding and
separating forces in
the nucleus. A proton
and a neutron can
proton proton form a stable nucleus
whereas two protons
can not.
Liquid drop model
It is far beyond the ambition of this book to give
a comprehensive understanding of these
processes. However, some penetration of the
topic is necessary to get a general feeling of the
fundamental aspects of stability and nuclear
decay. We use a model describing the exchange
of nuclear forces that is referred to as the “liquid
drop model”. This model gives an empirical
formula of the binding energy B of the nucleus.
Figure 5 The range of
the nuclear force is
short. Only neighbors
are interacting.
We explain the model step by step.
B ≈ a1*A
The binding energy is in the first approximation
proportional to the number of nucleons in the
nucleus. This is because the nuclear force has a
very short range. Only neighbors are affected
and can bind to each other. The nucleons in the
central part of the nucleus all have the same
number of neighbors. In a large nucleus the total
binding energy, B, is then approximately equal
to a constant*A where A is the number of
nucleons. The same is true between molecules
in a liquid, thus the model’s name.
B ≈ a1*A - a2*A2/3
Nucleons at the surface have fewer neighbors well. A nucleus with an odd number of protons
and binding sites and are more loosely bound. or neutrons is less stable than a nucleus with an
The total binding energy is decreased by a factor even number of both protons and neutrons. The
that is proportional to the surface of the nucleus. most unstable nuclei have an odd number of
The number of nucleons, A, is proportional to both protons and neutrons, e.g. 14N (Z=7, N=7).
the volume of the nucleus. Thus, the nucleus In fact, only seven odd-odd stable nuclides
radius is proportional to A1/3 and the surface to exist, all of them among the light elements. This
A2/3. last factor a5 takes care of this effect in the
following way
B ≈ a1*A - a2*A2/3 - a3*Z2/A1/3
apair A-3/4 for even-even nuclei
When the numbers of protons start to increase, a5 = 0 for even-odd nuclei
the Coulomb repulsing force starts to become -apair A-3/4 for odd-odd nuclei
more important. The Coulomb force is the sum
When this model is fitted to experimental data
of all proton combinations, F =Σ(Zi*Zj)/(rij) 2. If
the following parameters are obtained
we integrate this function as a function of
distance, we obtain a measure of the energy a1=15.76 MeV
related to the force. It can then be shown that a2=17.81 MeV
the negative contribution to the binding energy a3 =0.7105 MeV
is proportional to Z2/A1/3 a4 =94.81 MeV
apair = 34 MeV
B ≈ a1*A - a2*A2/3 - a3*Z2/A1/3 - a4*(A/2-Z)2/A
An interesting parameter is the mean binding
Nature seems to prefer to have about the same
per nucleon, B/A.
number of protons and neutrons in the nucleus.
This is seen in Figure 1 where for light nuclides
the nuclide chart is very close to a straight line
(equal number of protons and neutrons). For
heavier elements there seems to be a tendency
towards an excess of neutrons. This is
understandable if one considers the Coulomb
force contribution. Adding neutrons will
increase the distances between the protons,
which will lower the Coulomb force and total
binding energy in the nucleus increases. The
expression a4*(A/2-Z)2/A is only empirically
found but gives the wanted relations. If Z=A/2
i.e. the number of protons and neutrons is equal
the binding energy is at a maximum. The
contribution of this term is also less important
for heavier nuclides, which agrees with
experimental findings.

B ≈ a1*A - a2*A2/3- a3*Z2/A1/3- a4*(A/2-

Z)2/A+a5
Paired electrons in the outer electron shell give
chemically stable compounds, whereas unpaired
electrons give radicals. The most chemically
inert atoms are the noble gases (He, Ne, Ar, Kr,
Xe, Ra), which have closed electron shells. The
same phenomenon is found in the nucleus as
Figure 6 Mean binding energy per nucleon as a
function of mass number A. The liquid drop
model is fitted to measured data. The area from
0<A<50 is enlarged in the lower part of the
figure. As seen, the crude empirical model fits
the data well except for the light elements.

10
This value, which gives the mean energy needed
to separate one nucleon from the nucleus, varies
from a few MeV to about 9 MeV as illustrated
in Figure 6. The mean binding energy per
nucleon is a smoothly varying curve that has a
maximum around the iron part (A ≈ 60). If two
light elements are fused together they gain
energy. This effect is used in the fusion
principle to obtain nuclear power. If one heavy
element is split in two, the total binding energy
increases as well. The excess in energy is then
released and used in the fission principle to
obtain nuclear power.
As seen in Figure 6, the liquid drop model fits
experimental data quite well. However, at a
closer look, especially for the light elements
there are some disagreements. This is because
the nucleus can have closed proton and neutron
shells that are especially stable structures.
Critical numbers are 2, 8, 20, 28, 50, 82 and
126. This should be interpreted as if the nucleus
has eight protons (oxygen) there is a closed
proton shell and 20 neutrons will give a closed
neutron shell, etc. Of special interest are nuclei
with both a closed proton shell and a closed
neutron shell e.g. 16O (Z=8, N=8) or 40Ca
(Z=20, N=20). It has long been known that
some nuclides (so called magic nuclides) were
especially stable, but not the reason why. In the
close-up of Figure 6, these nuclides seem to
have extra high binding energy in comparison
with the surrounding nuclides.
RADIATION FROM RADIOACTIVE
NUCLIDES
Since radioactive material is the source of
radiation commonly encountered in a laboratory
(labeled compounds, calibration preparations),
radioactive decay is reviewed fairly thoroughly
in this section.
The concepts of nuclide and isotope are not
related with whether the nucleus is stable or
unstable. There are stable (non-radioactive)
nuclides and isotopes, just as there are unstable
(radioactive) nuclides and isotopes.
The number of neutrons and protons in the
stable nucleus obeys certain laws of symmetry.
11
The carbon isotope 12C is stable. The element
carbon has six protons (Z = 6) and the same
number of neutrons (N = Z = 6). If one neutron
is removed, the carbon isotope 11C (Z=6, N= 5)
is obtained. This nuclide is not stable and will
sooner or later undergo restructuring. If a
neutron is added, 13C (Z=6, N = 7) is obtained,
which is stable. However, its natural abundance
(1.1 %) is low compared with 12C.
Further addition of one neutron gives 14C(Z=6,
N=8), which also is radioactive. It is a general
rule (with a lot of exceptions) that the further
you move from the stable combination of
neutrons and protons, the more unstable the
nucleus becomes, which means that
restructuring to a stable condition occurs more
quickly (shorter half-life).
Radioactivity and decay time
The radioactive process, i. e. the decay of a
nucleus, is a statistical process. This means that
the decay time of an individual radioactive
nucleus can vary. But, for a large number of
radioactive atoms (N) the same fraction (dN)
always decays during the same time (dt). We
can write this mathematically as
dN = -λ N dt
where λ is called the decay constant and is the
probability for a nucleus to decay per time unit.
We can integrate this equation to obtain
N = No e- λ t
where No is the number of radioactive atoms at
t=0.
The number of decay per time unit (A = - dN/dt)
is called radioactivity. The relation between the
radioactivity and the number of radioactive
atoms is then A = λ N. Replacing this in the
formula above gives
A = Ao e- λ t
where Ao is the radioactivity at t=0. Ao is also
equal to λNo.
Every radioactive decay is associated with a
characteristic decay time. A common way to
express this is to use the half-life (T½) i.e. the
time it takes to half the number of radioactive
atoms. If we insert this time in the decay
formula, we get the following relation between
the half-life and the decay constant
Ao/2 = Ao e- λ T½ or
The conversion factor from the old unit to the
new unit is
1 Ci = 3.7 * 1010 Bq = 37 GBq (G = giga)
Table 3 Conversion from Curie (Ci) to
Becquerel (Bq) and the prefixes in the SI-system

T½ = ln(2)/ λ Prefix Ci Bq Prefix

6 16 37 PBq P=peta
M=mega MCi 10 3.7*10
We can also write the decay formula as k = kilo kCi 103 3.7*1013 37 TBq T=tera

A = Ao 2-t/T½
Ci 1 3.7*1010 37 GBq G=giga
m = milli mCi 10-3 3.7*107 37 MBq M=mega
After one half-life the radioactivity is reduced to µ=micro µCi 10-6 3.7*104 37 kBq k = kilo
a half, after two half-lives to a quarter, after n = nano nCi 10-9 3.7*101 37 Bq
three half-lives to an eight, and after four half- p = pico pCi 10-12 3.7*10-2 37 mBq m = milli
lives to a sixteenth. Since 37 µBq µ=micro
f = femto fCi 10-15 3.7*10-5
( 1/2 )10 = 1/1024 ~ 10-3
a = atto aCi 10-18 3.7*10-8 37 nBq n = nano

one may use as a rule of thumb that ten half-
lives reduces the radioactivity by a factor
thousand and 20 half-lives by a factor million of
its initial value.
The half-life varies considerably for the various
radioactive nuclides. It can be fractions of
seconds, to hundreds of years or more. We
usually express the half-life in seconds (s),
minutes (m), hours (h), days (d) and years (a).
Note that a is the SI-unit for the mean year =
365.24220 days.
The unit of radioactivity in the SI-system is the
Becquerel, after the French physicist who in
1896 discovered natural radioactivity. The unit
is abbreviated to Bq.
1 Bq = 1 decay / s
Decay per second is also written as dps, after
the initial letters of the English words. An
alternative way to express radioactivity is dpm,
decays per minute.
Surface radioactivity is given in Bq /m2,
radioactivity concentration in Bq /m3 and
specific radioactivity in Bq /kg.
Radioactive decay
Unstable nuclides have a surplus of energy
compared with the surrounding nuclides. By
making a transformation within the nucleus, this
surplus energy decreases and the remaining
nucleus will be more stable (the binding energy
increases). There are many ways to rearrange
the nucleus to obtain a more stable nucleus. As
an example, if we remove one neutron from e.g.
14
C we obtain the stable 13C. However, this is
usually not a spontaneous process. As seen in
Figure 6, one needs 5-8 MeV to remove one
nucleon. Even if the nucleus gains energy in the
end it does not have access to the primary
energy needed to run this process. For very
heavy nuclides the binding energy of the
neutrons decreases and here we can find
spontaneous neutron emission. However, in
general this process is rare.

Curie (Ci) is an old unit for radioactivity that is

still used. From the beginning it was defined as
the radioactivity of 1 g radium, but was later
standardized to 3.7 * 1010 decays per second.

12

The β - decay
A process that can occur spontaneously is the
beta-decay. The number of nucleons is kept
constant in the decay but a neutron is changed
into a proton, or vice versa. A particle of
positive (positron or β+ ) or negative charge
(negatron or β− ) is removed from the nucleus
along with energy (the β particle's rest mass +
kinetic energy).
The β particle has the same weight and the
same particle properties as an electron. In fact, it
can not, when it has lost its kinetic energy, be
distinguished from an electron. Still, we name it
a beta particle to mark that it was created in a
nuclear process in the nucleus. We will below
see that also electrons can be emitted in
radioactive decays but they all will come from
the electron shells around the nucleus.
In a radioactive nucleus rich in neutrons, the
decay occurs as if a neutron (n) were converted
to a proton (p):
n --> p + β− + energy (1a)
This reaction formula fulfils some criteria. On
both sides
1. the mass is about equal since the neutron
weight is the sum of the proton and the
electron weight
2. the charge is the same
3. the numbers of nucleons are the same
However, although the energy released per
decay is a constant it was found that the kinetic
energy of the beta particles varied between zero
and the decay energy. To explain the missing
energy the physicist Wolfgang Pauli 1930
suggested an additional particle now known as
the neutrino. This uncharged particle with little
or no mass was also emitted in the decay and
shared the decay energy with the beta particle.
25 years later the neutrino was experimentally
verified. The complete reaction formula for the
beta decay can then be written as
n --> p + β− + ν + energy (1b)
13
where ν is the neutrino. The bar above the
ν indicates that the neutrino is an antiparticle
(antineutrino).
Symmetry is an important feature of nature. For
most particles, there is an associated antiparticle
which has the same mass. Other properties like
charge will be the opposite. Charged particle-
antiparticle pairs can annihilate, i.e. the masses
will disappear and the released energy will be
converted to photons.
Formula 1b also fulfils another rule in particle
physics that you can not create particles out of
nothing. The number of heavy particles (the
nucleons) is the same on both sides. The same is
true for light particles like the electron and the
neutrino. Since particles and antiparticle cancel
out the number of light particle is then also the
same on both sides (zero).
The decay of 14C is an example of beta minus
decay.
14
C(Z=6, N=8) -->14N(Z=7, N=7)+β−+ ν +energy (2}
Figure 7 Decay scheme for 14C. The x-axis
gives the number of protons in the nucleus and
the y-axis its energy content. By converting a
neutron to a proton, the number of protons in
the nucleus increases and at the same time it
becomes more stable. In the process, a
negatively charged beta particle is emitted.
The β+ decay can be regarded as a proton that is
converted to a neutron.
p --> n + β+ + ν + energy (3)
The β+ is now the antiparticle and cancel out the
“real” neutrino to sum up light particles on the
right side to be zero.
An example of the positron decay is:
11
C(Z=6, N=5) -->11B(Z=5, N=6) +β+ +ν +energy (4}
11C
Binding energy
Z=6, N=5
∆E=2 MeV
11B
Z=5, N=6
5 6
Number of protons in the nucleus
Figure 8 Decay scheme for 11C. The x-axis
gives the number of protons in the nucleus and
the y-axis its energy content. By converting a
proton to a neutron, the number of protons in
the nucleus decreases and at the same time it
becomes more stable. In the process, a
positively charged beta particle is emitted.
The radioactive nuclides used in laboratory
work usually have an excess of neutrons and
consequently emit their energy in the form a β-
particle, which is ejected from the nucleus with
a specific kinetic energy and decelerates in the
surrounding material.
The kinetic energy of the emitted particles is
determined by the energy released in
transformation of the nucleus. This may be
illustrated in an energy diagram or a decay
scheme. Figure 7 shows such a diagram for 14C
and Figure 8 for 11C.
The 14C-decay releases energy of 156 keV and
the beta particle has a maximum kinetic energy
of 156 keV. The 11C decay releases 2 MeV but
the maximum energy of the β+ is only about 1
MeV. This is because two new electron masses
are created in the β+ decay, which corresponds
to 1 MeV. We can understand this by looking
14
at the mass balance in equations (1b) and (3).
The mass of the neutrino can be neglected.
The mass of a neutron (Mn) is approximately
equal to the mass of a proton (Mp) and a beta
particle (Mβ). Equation (1b) is then in a mass
balance
Mn ≅ Mp + Mβ
In equation (3) we have the mass of a proton on
the left side and the mass of a neutron and a
beta particle on the right side. The only way to
get the equation in balance is to add two beta
masses on the left side.
2* Mβ + Mp = Mn + Mβ
According to the Einstein formula E = Mc2. The
mass of two beta particles corresponds to 1,022
MeV. In the 11C decay, a total energy release of
2 MeV is available. Of this energy about 1 MeV
is taken to account for the extra mass created in
the decay, leaving 1 MeV to be shared by the
positron and the neutrino. In the decay scheme
in Figure 8, this is indicated by the broken
arrow from 11C to 11B. The first vertical part
indicates the energy needed within the 11C
nucleus to create the two electron masses. The
diagonal part of arrow indicates the part of the
decay energy that is converted into kinetic
energy of emitted particles.
The decay energy
The decay of 14C always releases 156 keV.
Other beta decays, i.e. 3H and 32P (see Table 1),
are also associated with a unique energy release.
Table 1 Decay parameters of some common
radionuclides.
Nuclide Half -life Residual Decay energy, Eo
T½ KeV
3
H 12.26 a 3
He 18
14
C 5 760 a 14
N 156
32 32
P 14.3 days S 1710
The β-particles share this energy with the
neutrino randomly. The sum of the β−particle
and the neutrino kinetic energies are then equal Q-value
to the decay energy, Eo:
Each nuclear transition is characterized by a
Eβ + Eν = Eo fixed energy release or a Q-value. If the Q-value
is positive, surplus energy is emitted in the
In most beta decay the relative energy transformation. If the Q-value is negative,
distribution of the β-particles has the same additional energy is needed to enable the
shape (Figure 9). transformation. A spontaneous radioactive
decay has always a positive Q-value, whereas a
Relative number nuclear reaction in e.g. a nuclear reactor often is
of β -particles associated with a negative Q-value. The energy
released in a nuclear decay can be used to create
new mass or to give emitted particles kinetic
energy.

Pure beta decays go from the ground level of the

radioactive nucleus to the ground level in the
daughter nuclide. Three bodies share the decay
energy, the beta particle, the neutrino and the
recoiling rest nucleus. However, since the mass
of the recoiling nucleus is much larger than for
Figure 9 Energy spectrum of β −particles at the other particles, the energy of the recoil
radioactive β-decay. The energy is given on the nucleus is virtually zero. The total reaction
x-axis and the number of particles per energy energy is then statistically distributed between
bin on the y-axis. the beta and the neutrino, and the Q-value is
numerically close to (Eβ)max .
The distribution of energy to the particles is
determined statistics. The neutrino is an More generally, the decay may produce excited
uncharged particle of unknown, but very small, states in the daughter nuclide. The nucleons, i.e.
mass. It seldom interacts with matter and its the protons and the neutrons, create quantified
energy can in practice be ignored. Only the states of the nucleus. In the ground state,
β−energy, Eβ, can be transferred to the movements and energy contents are at a
surrounding material. The maximal beta energy minimum. In an excited or deformed state, the
and the mean beta energy can be written as: energy content is higher. The nucleus can
suddenly change its state, from a more excited
(Eβ)max = Eo E β ≈ Eo/3 state to a less excited or deformed state. In this
process electromagnetic photons, i.e. gamma
When calculating the maximum range of the rays, are emitted. The gamma rays carry away
β −particles, (Εβ)max is used. parts of the total decay energy. For each photon
(gamma quantum) emitted, the excitation
energy of the nucleus diminishes until it finally
Table 2 Maximum range of β -particles in air reaches the ground energy state.
and water.
Nuclide (Eβ)max Range (mm) An example is the decay of 60Co what can be
illustrated in the decay scheme below (Figure
(keV) Air Water
10). It is a beta minus decay emitting a beta
3H 18 6.5 0.007 particle with a maximal energy of 2.824 –
14C 159 300 0.3 2.506=0.318 MeV. The decay created an
excited daughter nucleus 60Ni. This excited
32P 1710 7700 7.9 levels emits a gamma of 2.506-1.332=1.173
MeV. A new excited level is created which also

15
emits a gamma of 1,332 Mev. If we ad up the
total energy emitted 0.318+1.173+1.332 we end
up with the Q-value of the decay (2.824 MeV).
E n e rg y
(M e V )
2 .8 2 4
2 .5 0 6
1 .1 7 3 M e V
gam m a
e m itte d
1 .3 3 2
1 .3 3 2 M e V
gam m a
e m itte d
0 in 93 % of all decays.
Figure 10 A simplified decay scheme for 60Co.
Only dominant radiation is included.
Electron capture
The total energy release (the Q-value) in a
positron decay has to be more than 1 MeV to
compensate for the increase in mass (see
above). An alternative route of decay that does
not need this extra energy is electron capture.
Instead of converting a proton to a neutron and
a positron, the nucleus can attract an electron
from the surrounding electron shells and join it
with a proton to create a neutron.
-
e + p --> n + ν
The effect is the same as in the positron decay; a
proton is converted to a neutron. However,
since no beta particle is emitted the full excess
energy in the decay of the nucleus has to be
given to the neutrino. Accordingly, in this type
of decay monoenergetic neutrino particles are
emitted. The decay of 125I is a typical example
of this process (Figure 11 and 12).
Although no beta particle is emitted the electron
capture is a variant of the beta decay since the
same physics is involved. There is then three
branches of the beta decay, the beta minus, the
beta plus and the electron capture.
Conversion electrons
The decay of 125I gives a new example of a
decay process ending in an excited state of the
daughter nuclide (Figure 11). As said above, the
16
common way to emit the energy in the excited
state is by emitting a gamma ray. This is also
done in the 125I decay, but only in 7 % per
decay. The excited state, 35.5 keV, is very close
to the binding energy of the K-shell electrons
(33 keV). There is a probability that a K-shell
electron is inside the nucleus due to the
quantum mechanical nature of the electron.
There is also a probability that the excited state
interacts with and transfers its energy to the K-
shell electron. The transferred energy is higher
than the binding energy, and we obtain a
monoenergetic electron, a conversion electron,
The excited state does not only interact with the
K-shell but also with the L-shells and the M-
shells although the probability is less (the
electrons are further away from the nucleus and
thus the probability of being inside the nucleus
is smaller). The energy of the conversion
electrons varies according with the formula
below
E = 35.5 keV - Eshell
where Eshell is the binding energy of the shell
from which the electron is emitted.
In the 60Co decay a small amount of conversion
electrons is found as well. In fact, an excited
state in a nucleus always has two ways to de-
excite, either by emitting a gamma or by
converting the excited energy to a shell electron.
The heavier the nuclide is, the higher the
percentage of conversion electrons. A heavy
nucleus has many protons in the nucleus and a
strong attractive force on the shell electrons that
are close to the nucleus. Thus, the probability
that the excited state will interact with the
electrons increases.
Another important factor that affects the
probability of emitting conversion electrons is
the energy of the excited level. Energies close to
the binding energy of the shell electrons give
the highest probabilities.
The electron capture and the conversion
electrons both create holes in the electron shell.
The process to fill these holes created a cascade
of events and the emission of electrons called energy is transmitted as X-rays or free electrons
Auger electrons and characteristic X-rays. that are dislodged from the atom (Auger
These processes are exemplified and explained electrons). This process is seen in more detail in
in more details in the decay of 125I. Figure 12.

Decay of 125I
125
I is a neutron deficient nucleus (the stable Electron
Decay of excited
125Te to ground
isotope of iodine is 127I). It decays to 100% by capture, EC
state by internal
electron capture since its Q-value of 186 keV is conversion
far to low for positron decay. The decay scheme Step 1 Step 4
is seen in figure 11 and shows that the decay in
100% yields an excited level in the daughter
Cascade of
nuclide Tellurium-125. This excited level of characteristic Conversion
35.5 keV is just above the binding energy of the X-rays electron
k-shell electrons (33 keV). This excited level is and Auger
de-excited by gamma emission but only in 7% Step 2 Step 5
per decay. More likely (93% per decay) is that a
conversion electron (energy 35.5-33=2.5 keV)
125Te
is emitted. 125Te in an in
excited state ground state

125I (Z = 53, N = 72)

Step 3 Step 6

Figure 12 Details of the 125I-decay

149 keV
EC (Electron capture)
125Te in an excited stage
emitted gamma (7 %) or
35.5 keV
conversion electron (93 %)
125Te (Z = 52, N = 73)
Figure 11 Decay scheme of 125I. The number of
protons in the nucleus is on the x-axis and the
energy content of the nucleus on the y-axis. By
capturing an orbital electron, a proton is
converted to a neutron and at the same time, the
nucleus becomes more stable. In the process, a
neutrino is emitted and carries away the energy
which is formed. The new nucleus is formed in a
excited state, but after a time it is de-excited to
a more stable state. In this process, either a
gamma ray or a conversion electron is emitted.
Both at the electron capture, when an electron is
drawn into the nucleus, and at the forming of
conversion electrons, holes appear in the
electron shell around the nucleus. These holes
become filled by electrons from the outer shell.
In this process, the disparity in the binding
Hence, the main radiation from 125I are K shell
X-rays of 27 keV and Auger electrons of low
energy. The range of the electrons is
comparable to that of β − particles from 3H.
Generally, the photon radiation is brought to a
standstill practically by two millimeters of
brass, whereas several decimeters of water are
required.
Alpha decay
Alpha emitting radionuclides are not commonly
used in the laboratory and will only briefly be
treated here. The physics involved is different
from the beta decay and can actually be
described as a tunneling effect through the
potential barrier around the nucleus. The
following description is just to get an idea of
what this potential barrier is and the main
features of the decay.
Consider a free He++ ion outside a heavy
nucleus (see Figure 13). If you try to move the
He++ ion closer, an increasing Coulomb force

17
in-between occurs. This force tries to repel the
He++ ion until it is so close to the nucleus that
the strong force starts to act. The He++ ion is
then trapped inside the so-called nuclear
potential, which is created in a balance between
the nuclear force and the Coulomb force. The
barrier the He++ ion has to climb before it
reaches the attracting nuclear force is called the
coulomb barrier. Its height depends on the
number of protons in the nucleus, but is in the
order of 5-7 MeV for heavy nuclei.
When particles are trapped in the nucleus they
loose their identity. However, the He++ ions
consist of two protons and two neutrons. Both
the proton shell and the neutron shell are closed
and the He++ ion is a very stable combination of
nucleons. One might consider that this stable
combination can exist in the nuclear “soup” in
the form of a virtual α-particle. Also, these
virtual α particles may populate different energy
levels in the nucleus. In the nucleus, the energy
content is quantified and different quantified
energy levels exist.
In Figure 13, two such levels are indicated. In
the bound level 1, the virtual α particle cannot
escape the nucleus since it is bouncing against
an infinitely thick potential wall. In the bound
level 2, however, a potential wall exists that
keeps the virtual α particle trapped but it is not
infinitely thick. The α-particle may penetrate
the wall by, what is known in quantum
mechanics as, tunneling. Suddenly, the α-
particle appears in a materialized form outside
the Coulomb barrier with energy corresponding
to the excited level.
He++ repulsive
+ coulomb bound level 2
+ force
bound level 1
Nucleus Z>80
heavy positivly
charged
18
Figure 13 Schematic view of the alpha decay
The probability of decay is highly determined
by the thickness of the barrier, and the half-life
of useful radioactive sources varies from hours
to many thousands of years. Alpha particles
appear in one or more energy groups that are,
for all practical purposes, monoenergetic. For
each distinct transition between initial and final
nucleus, a fixed energy difference or Q-value
characterizes the decay. This energy is shared
between the alpha particle and the recoil
nucleus.
The alpha particle energy correlates strongly
with the half-life of the parent nuclide; the
highest energies have the shortest half-lives.
Above 7 MeV, the half-life can be expected to
be less a few hours and. On the other hand, if
the energy drops below 4 MeV, the barrier
penetration probability becomes very small and
the half-life of the isotope will be very large.
Probably the most commonly used source for
alpha particles is 241Am (T½=433 a) with an α-
energy of 5.5 MeV.
Because alpha particles lose energy rapidly in
materials, alpha particle sources must be
prepared in very thin layers. To contain the
radioactive material, typical sources are covered
with a metallic foil or other material that must
also be kept very thin if the original energy and
monoenergetic nature of the alpha emission is to
be preserved.
Almost all naturally occurring alpha emitters are
heavy elements with Z >83. The principle
features of alpha decay can be learned from the
example of 226Ra:
226 Ra 222 4

Rn + He
88 86 2
The energy released (Q-value = 4.88 MeV) in
the decay arises from a net loss in the masses
MRa, MRn and MHe of the radium, radon, and
helium nuclei:
Q = MRa - MRn- MHe
The general equation for the energy release in
the alpha decay is then
Qα=Mparent-Mdaughter-MHe
Spontaneous fission
The fission process splits the nucleus into two
parts of almost equal size. In the process free
neutrons are also emitted. Fission fragments are
used to calibrate and test detectors intended for
general application in heavy ion measurements.
The most widely used example is 252Cf, which
undergoes spontaneous fission (3% per decay)
and alpha decay (97% per decay) with a half-life
of 2.65.
Each fission gives rise to two fission fragments,
which, by the conservation of momentum, are
emitted in opposite directions. Because the
normal physical form for a spontaneous fission
source is a thin deposit on a flat backing, only
one fragment per fission can escape from the
surface, whereas the other is lost by absorption
within the backing. The spontaneous fission of
252
Cf also liberates a number of fast neutrons.
Natural radioactivity
Some heavy elements (232Th, 235U and 238U) has
such long half-lives (109-1010 a) that they have
survived the time since the where created in the
Big Bang and are still to be found in appreciable
amounts. In there decays they produce a series
of more short-lived radioactive daughters. Some
of these daughters we are very much aware of
e.g. the alpha emitter Radon-222, which is a
noble gas and that can penetrate into our
house
40K
1461 keV
EC (11 %)
(T½=1.28 109 a)
β- (89 %)
1312 keV
β+ (0.001 %)
482 keV
40Ar
Figure 14 Decay scheme for 40K
Few lighter elements have naturally occurring
radioisotopes with that long half-live. The most
important from the standpoint of human
exposure is 40K, which is about 0.01% abundant
and has a half-life of 1.26 x 109 years (Figure
14). The nuclide can decay by β- emission
(89%), EC (11%), or β+ emission (0.001%).
The maximum β- energy is 1.314 MeV. This
isotope is an important source of human internal
radiation exposure, because potassium is a
natural constituent of plants and animals.
Another important, naturally occurring
radioisotope is 14C, used in carbon dating. It is
produced by the bombardment of nitrogen in the
atmosphere by cosmic rays, causing the reaction
14
N(n, p)14C. Its half-life is 5730 yr. The
radioisotope, existing as CO2 in the atmosphere,
is used by e.g. trees and becomes fixed in their
structure through photosynthesis. When an tree
dies the incorporation of new 14C atoms stop
and the number will start to decrease due to the
radioactive decay. By, in a sample, comparing
the number of 14C with the number of stable 12C
one can then calculate the time that has passed
since the tree was cut and used for e.g. building
a house. Thus, the age of such objects can be
determined. The atmospheric testing of nuclear
weapons has significantly added to the world's
inventory of 14C.
DATA ON RADIOACTIVE NUCLIDES
There are several books available, with all
relevant information on decay-time, type and
number of particles emitted per decay, energy
per particle, etc. However, the fastest and
cheapest way to obtain information today is by
the Internet. One good site is
http://www.nndc.bnl.gov/nudat2/indx_dec.jsp
40Ca
Below there is an example of the 125I decay and
with some explanations.
19
20
 CHAPTER 3
INTERACTION WITH MATTER
energy exceeds the binding energy and we
Matter is composed by a number of different obtain a free electron, ionization. The same
atoms in chemical combinations. A piece of thing is true for an electron or a beta particle
iron is in a macroscopic sense solid and difficult that will experience a force of the same strength
to penetrate. However, at the dimensions of but repulsive.
ionization radiation, matter is rather empty and
without any major mechanical obstructions to
prevent radiation to pass.

Consider a neutron entering a piece of iron. The

iron atom has a dimension of 10-10 meters but
the iron nucleus is only 10-15 meters, and the
neutron itself is even smaller. The electrons
have no known extension and since the neutron
has no charge it will not feel any electric forces.
The neutron will probably pass through a thin
iron foil without noticing the material.
However, if the foil grows thicker it will
increase the likelihood that the neutron will hit
one of the atom nuclei, cause a nuclear reaction
and transfer its energy to the matter.

If we exchange the neutron for a proton, a

dramatic change occurs. The proton will
experience that matter is full of charge. Each
orbit electron in the vicinity of the proton will
feel a force of attraction, the Coulomb force
(Figure 1).
Figure 2 An α-particle transfers so much
energy that an ionization occurs. Since the α-
particle and the orbital electron have opposite
polarity, there is an attractive in-between.

electron
mass m
charge -q

distance r
closest
distance d
F = q*q/r 2

proton
mass M
velocity v
charge +q

Figure 1 A proton that enters into matter

experiences an attractive force from all the
orbit electrons in the vicinity.
We can say that we have a collision between
two electrical fields and in which energy is
transferred to the orbit electron. Sometimes the
Figure 3 The electrical field of a beta particle
collides with that of an outer orbit electron. So
much energy is transferred that the orbital
electron is dislodged (ionization). Since the
incoming electron and the orbital electron have
same polarity, a repulsive force is created.

21
However, the electron and the proton differ
considerably; they have different masses and
different speeds. An electron with the energy of
1 MeV has a speed close to the speed of light
(3*108 m/s). The time it will spend in the
vicinity of an atom and an orbit electron will be
10-18 - 10-19 seconds. The force will act during a
very short time and the impulse ∫F*dt
transferred from the incoming electron to the
orbit electron will be small. A proton of the
same energy (1 MeV) will have a speed 20
times lower than the electron. The time spent in
the vicinity of the orbit electrons will be
correspondingly longer and the transferred
impulse larger. From this simple consideration
we can draw two important conclusions that are
true for charged particles:
1. The heavier the particle is, the more energy
is transferred per collision.
2. The lower the kinetic energy of the particle
is, the more energy is transferred.
The expression "ionizing radiation" includes
many types of "radiation". The most important
kinds of radiation are classified according to
how they transfer energy to matter in the
diagram below (Figure 4).
IONIZING RADIATION
Electro-magnetic Particles
radiation, photons (protons, electrons,
(X-rays and gamma) alpha and neutrons)
Uncharged particles Charged particles
(neutrons) (protons, electrones
and alpha)
Light, charged Heavy, charged
particles particles
(electrones) (protons and alpha)
Figure 4 Schematic subdivision of the different
types of ionizing radiation.
The direction of heavy, charged particles is
scarcely affected by collisions with orbital
electrons. They continue to travel mainly
22
straightforward to the end of their range,
deviating only slightly from a straight line.
Thus, heavy, charged particles have a well-
defined range and monoenergetic particles
travel about the same distance in a given
material.
When heavy, charged particles are obstructed
their speed diminishes. Consequently, they
spend a longer time close to the next orbital
electron and transfer more energy to it. Then,
the number of ion pairs formed per unit of
length increases (see Figure 5).
Figure 5 Deceleration of an α −particle in air.
The α -particle at high energy has high speed.
The time it spends near each orbit electron is
short; hence, the transferred energy is small
and the probability for ionization is relatively
small. When the energy diminishes so does the
velocity. The probability of ionization increases
and a typical ionization peak at the end of the
particle track (the Bragg peak) occurs.
If the incoming charged particle is an electron,
the rest mass of this particle is the same as for
the orbital electron. When two particles with the
same mass collide, a great deal of energy
(almost all) can be transferred in a direct impact
and the incoming particle will come to a
complete stop. For two electrons we have a
problem; they are identical. We cannot with any
means know which one is the incoming particle
and which one is the orbital electron. For
practical reasons, therefore, we define the
electron with the highest energy after the
collision to be the incoming particle.
At each collision, light charged particles can Bremsstrahlung
change direction very markedly and by repeated
large directional changes can even be scattered When electrical charges are accelerated or
backwards to the direction of their origin decelerated, they can emit electromagnetic
(Figure 6). Hence, the range of electrons differs radiation. We call this roentgen, continuous X-
considerably. However, a maximum range can rays or bremsstrahlung. Synchrotron light is of
be given (Figure 7). the same nature.

At these collisions, the energy that is transferred

is frequently sufficient to ionize an orbital
electron. The energy might even be so high that
the free electron, in turn, can cause new
ionization. Hence, part of the energy of the
primary charged particles is deposited at some
distance from the original track of the particle.
Secondarily formed electrons, δ−particles, can
give rise to further ionization tracks.

Back-scattered Figure 8a An incoming electron feels the

electron
Coulomb force from the nucleus and is
deflected. The electron speed and kinetic energy
Bremsstrahlung
2 0 4 0 6 0 8 0 1 0 0
S e r i e s 1
decreases in the process - the electron
decelerates. The difference in kinetic energy
between in-going and out-going electron is
emitted as an X-ray photon.

10
Coulomb collision
Energy loss (MeV/g*cm2)

Figure 6 Electrons, which penetrate material,

collide with orbit electrons of the same mass as
the incoming particles. They can effect marked 1.0
directional changes including scattering in the
Water
direction opposite to the original course. Lead

0.1

Bremsstrahlung
0.01
0.1 1.0 10 100
Electron energy (MeV)
Figure 8b Loss of energy by electrons per track
length (expressed in mass per unit area) as a
function of electron energy. A heavy and a light
material show how the atomic number of the
material influences the two ways by which
Figure 7 A schematic figure illustrating the electrons transfer energy to material.
range of monoenergetic electrons. Most of the
electrons are absorbed within a distance
Heavy particles lose only little energy per
shorter than the maximal range.
collision and are only decelerated in small steps.

23
Thus, for heavy particles the loss by
bremsstrahlung is of little importance.
Electrons, on the other hand, often lose a great
deal of their energy by deceleration in the strong
fields of heavy atoms; hence, the loss by
bremsstrahlung is of more importance. The
result of continuous radiation increases with the
electron energy and atomic number of the
absorber. For high electron energies and heavy
elements (e.g. lead), the highest X-rays yield is
obtained (Figure 8b).
Stopping power
Charged particles gradually lose their energy
(they are decelerated) by many close
"collisions" with orbital electrons in the
irradiated material. The ability of different
materials to decelerate charged particles varies
considerably depending on the charge and
energy of the charged particles, and the density
and atomic number of the irradiated material.
The deceleration capacity attributed to a given
material is denoted by how much energy (∆Ε) is
expended in a given length (∆l) of the charged
particle’s track.
Stopping power = (∆Ε) / (∆l)
STOPPING POWER AS A FUNCTION OF
PARTICLE ENERGY
10000
Stopping power (MeV/cm)
1000
100
10
1 The radius r can vary, which gives different
0.01 0.1
Particle energy (MeV)
Figure 9 Stopping power in water for electrons,
protons and alpha particles as a function of
energy.
Stopping power is a parameter that describes the
material’s ability to stop charged radiation. For
different combinations of particles and material,
the ability to stop the radiation can be calculated
and tabulated. Stopping power data for electrons
in lead and water are given in Figure 8.
Stopping power for water is given for electrons,
protons and α- particles in Figure 9.
Linear energy transfer, LET
The energy that charged particles lose by
deceleration is transferred to the irradiated
material in different ways. The energy can be
deposited more or less concentrated around the
primary particles' track. Direct ionization results
in energy deposition in the vicinity of the
particles' track. However, secondary particles,
such as bremsstrahlung and neutrons (in nuclear
reactions) with no charge, may deposit their
energy far away from the particle track. To
understand the effect of radiation on living
material, it is also important to have a measure
of how densely the radiation's energy is
deposited around the particles' track. Hence, the
concept Linear Energy Transfer (LET) was
introduced. It denotes the amount of energy the
specified particle emits per unit track length and
that is distributed near it.
DNA double helix
cylinder diameter 2r
5.3 MeV α - track
Figure 10 Illustration of the ionization pattern
around an α-particle track. Each black spot
electron represent one ionization. Around the track, the
proton
alfa
ionization density is highest. The limited
stopping power or the linear energy transfer,
LETr, is given as the energy per track length
deposited inside a cylinder with the radius r.
1 10
LET-values. Often, the radius r is set to be the
diameter of the DNA double helix. Instead of
giving the radius length in nanometers, it is
given as the energy of an electron with the
range r.
When living tissue is irradiated, LET usually
denotes the energy absorbed per unit length
around the particles' track within a cylinder. The
diameter of this cylinder is chosen to be in the
24
same order as a biological critical structure, e.g.
the cell nucleus or the DNA-molecule.
High LET and low LET
It is of interest to understand how many
ionization are created in the DNA molecule of
different types of radiation. A rough estimate is
made in the following.
The stopping power for electrons, protons and
alphas (see Figure 9) is recalculated and
expressed in number of ionization/nm (Figure
11). It is seen that for electrons of all energies
the number is below 0.1 ionization per nm. The
more correct use of LET, not stopping power,
gives an even lower number. Since the DNA
molecule has a diameter of 2 nm, this indicates
that it is very unlikely that electrons of any
energy will cause two ionizations so close that a
double strand break occurs.
High energy protons (E > 10 MeV) have the
same pattern but low energy protons with
energies <1 MeV yield more than 1
ionization/nm. Alpha particles over a wide
range of energies also yield more than 1
ionization/nm. If the probability of causing a
double strand break is low, we have low LET
radiation whereas if the probability is high, high
LET.
IONIZATION DENSITY
10
1 Energy h* ν
0.01 0.1 1 10
Ions/nm
0.1
0.01
0.001
0.01 0.1 1
Particle energy MeV
10
Figure 11 Stopping power in water given as
number of ionizations per nanometer for
electrons, protons and alpha particles and as a
function of particle energy.
Photon interactions
Non-charged particles such as photons and
neutrons do not feel the Coulomb force and
cannot transfer their energy to matter this way.
However, in occasional interactions they may
transfer all or much of their energy to charged
particles in the irradiated material. Thus, these
secondary charged particles (photons give rise
to secondary electrons and neutrons chiefly to
secondary protons) can continue to ionize
through the charge interaction. Since the
ionization of the primary interactions is small
compared with the ionization of the secondary
radiation, we sometimes refer to photons and
neutrons as indirect ionization radiation. Only
the interaction of photons is described here.
Regarding the interaction of photons with
matter, three processes can be distinguished.
The photoelectric effect implies that a bound
electron, mainly a K-shell electron, absorbs the
photon in one of the innermost atom shells. This
electron, the photoelectron, obtains a kinetic
energy that corresponds to the photon energy
minus the electron's binding energy in the shell.
An electron from one of the outer shells then
fills the “hole” in the electron shell, and
characteristic X-rays (or Auger electrons) are
emitted. The energy balance in the transfer is
Auger electron
Characteristic
X-ray
Incoming photon
Photo electron
electron
proton
Figure 12 The photoelectric effect. A photon
alfa interacts with a K-shell electron. All the
photon's energy is used to displace the electron
and give it kinetic energy. When the "hole" of
the electron is filled with an electron from an
outer shell, energy is liberated as a photon
(characteristic X-radiation) or is transferred to
another orbit electron (Auger electron) that
leaves the atom.
hν = Εel + Βe
25
where hν is the photon energy, Eel is the
electron's kinetic energy and Be is the binding
energy.
The photoelectric effect is not significant in air
and soft tissue (which includes material of low
atomic number) for photon energies over 200
keV.
In the Compton effect, the electron binding
energy is always insignificant in relation to the
photon energy (Figure 13). The Compton effect
can be interpreted as an impact process in which
the photons collide with a free electron.
Figure 13 Compton effect. A photon interacts
with an electron in an outer orbit. Only part of
the photon's energy is used to dislodge the
electron and deliver kinetic energy. The
remaining energy is transmitted to a new
photon that has a different direction and less
energy than the incoming photon.
Hence, in this process the main electrons
involved are the outer, loosely bound electrons.
The physical laws of impact apply, since the
system's total energy and momentum are the
same before as after impact. This results in a
photon with lower energy and indeed greater
wavelength. The energy and the angle between
the scattered photons and electrons can be
calculated by using the laws on conservation of
energy and impulse.
The probability of “Compton scattering" per
electron of per unit mass is about the same for
all material, but it decreases somewhat with
increasing photon energy. The Compton effect
is significant for photon energies between 100
keV and 10 MeV. Part of the incoming photon
26
energy, which is transmitted to the Compton
electrons, is absorbed within a tiny volume
around the region where the process occurs.
The amount of energy transmitted to the
scattered photons, however, is usually not
absorbed in the immediate vicinity. The photon
can cover a considerable distance before being
absorbed.
The pair production effect (Figure 14) signifies
that the photon interacts with the electrical field
in the neighborhood of an atom's nucleus. The
photon disappears and its energy is converted to
an electron and a positron (see Figure 15). The
electron mass is equivalent to 0.511 MeV. Since
two electron masses are created the incoming
photon must have energy of at least 1.02 MeV.
Surplus energy is given as kinetic energy to the
electrons.
Incoming photon e-
Energy h*ν
Recoil
e+
Figure 14 Pair production. The incoming
photon energy is so great that it can create two
new particles, an electron and a proton. The
entire energy of the incoming photon is
required to create the mass of these particles
(2*511 keV) and give these particles their
kinetic energy.
A positron can be said to be a positive electron.
It has the same mass as an electron but a
positive, not a negative unit charge. On
decelerating in matter, the positron behaves in
the same way as the electron except that the
positron at the end of the track attracts an
electron and is "annihilated" by it. This means
that the energy that is equivalent to two electron
rest masses (2*511 keV) is converted to two
511 keV photons which are emitted in two
directly opposite directions. This radiation is
called annihilation radiation.
Pair production and annihilation are thus two
diametrically opposite processes. In pair
production, a photon is lost and two light
particles are formed, a negatron (ordinary
electron) and a positron. Both of these particles
are decelerated whereupon the positron is
annihilated by an orbital electron during the
emission of two 511 keV photons.
100
Photoelectric Pair
80 effect production
Atomic number
60
40
Compton
20 effect
0
0.01 0.1 1 10
Photon energy (MeV)
Figure 15 Dependent on the energy of the
photons and the atomic number, the probability
of different photon interactions varies. In the
figure is shown a special case when the
probability for the Compton effect is equal to
the probability of the photo-effect (the curve to
the left) and when the probability for the
Compton effect is equal to the probability of
pair production (curve to the right).
The relative significance of the different
processes can be denoted as functions of photon
energy and absorbing material. The
combinations of energy and atomic number by
which the probability of obtaining the
photoelectric or the Compton effect are equally
large (Figure 15).
For water and soft tissue with "effective atomic
numbers" lower than 10, the photo-electric
effect dominates for photon energies < 100
keV, the Compton effect for > 100 keV and
< 10 MeV, and pair production for photon
energies > 10 MeV.
For low photon energies and for high Z material
where the photoelectric effect dominates,
absorption discontinuities occur (Figure 16) at
photon energies equivalent to the binding
energies of the inner shell electron. Just below,
the photon energy is too low to interact and the
electrons in this shell do not contribute at all to
the attenuation processes. Just above, the
probability of photon absorption is very high.
105
Probability of the
photo effect (m-1)
104
103
102
0 100 200 300 400
Photon energy (keV)
100 Figure 16 The probability for the photo effect in
lead as a function of photon energy. Some
discreet jumps in the curve are seen at energies
corresponding to binding energies of the orbit
electrons.
Heavy elements, where the binding energy of
the K-shell electrons is high, are therefore likely
to interact with high-energy photons. For
example: In lead (atomic number = 82) the
photoelectric effect dominates for photons with
energies of < 0.5 MeV, the Compton effect at
photon energies > 0.5 MeV and < 5 MeV, and
pair conversion for photons with energies >
5MeV.
Attenuation of photons
For photons, as for neutrons, matter looks
empty. Photons can pass a long way through a
piece of material before they interact. They may
even penetrate a piece of matter without any
interaction. However, when the photons interact
they lose all or most of their energy, which is
transferred to charged particles.
The formula for the attenuation (damping) of
photon radiation in material is:
N = No e -µ x
where
27
N is the number of non-scattered photons that exponential attenuation and there is always a
leave the absorber certain finite probability that some photons will
No the number of photons incident on the penetrate even thick material. What we can
absorber and calculate is the fraction of photons that will
µ is the linear attenuating coefficient (m-1), interact in a certain thickness of irradiated
which is a measure of probability per unit material.
length for a photon to interact
x is the thickness of the absorber. The linear attenuation coefficient (µ) is the sum
of the photoelectric effect (τ), the Compton
effect (σ) and pair conversion effect (κ). The
photoelectric effect dominates at low photon
energies, the Compton effect for moderate
energies and the pair conversion effect for high
photon energies. The contributions of different
attenuating coefficients in aluminum are shown
in Figure 18.

The thickness of the absorber that decreases the

Figure 17 A narrow beam of photons, No, hit a number of incoming photons by a factor of two
piece of material with thickness x. Some is of special interest and has a special name, the
photons are absorbed or attenuated out of the half layer value (HLV). It is defined as
beam. The number of photons hitting the
detector is then N. 1/2 No = No e −µ HLV

104
It is related to the attenuation coefficient in the
following way
Probability for different photon

103
interaction effects (m-1)

HLV = ln 2/µ = 0.693/µ

102 µ=σ+τ+κ

So far, only the linear attenuating coefficient

10
has been mentioned. It is often more convenient
to deal with attenuation coefficient per unit
σ τ κ
1
σ
mass, which is given by µ divided by ρ, the
density of the material. This gives the mass
0.1 attenuation coefficient, which is also denoted by
0.1 1 10 100 1000
Photon energy (MeV) µ/ρ.

Figure 18 The probability in aluminum for the When using the mass attenuation coefficient per
three main ways in which photons interact as a unit mass, it is necessary to multiply by both the
function of photon energy. The different effects thickness of the material and its density to
are denoted. obtain a non-dimensional exponent in the
τ = photo effect expression.
σ = the Compton effect
N = No e - µ/ρ ∗ ρ · x
κ = the pair production effect
µ = the sum of the three separate effects gives
where
the total attenuation coefficient.
µ has the dimension m-1, ρ kg/m3. (µ/ρ) has the
Unlike charged particles, photon radiation has
dimension m2/kg and x m, hence it follows that
no fixed range or maximum range. The
the exponent has no dimension. Variations of
interaction of photon radiation leads to an

28
the mass attenuation coefficient for lead,
copper, water and air are illustrated in Figure
19.

lead
Mass attenuation coefficient (m2/kg)

10-1

copper

water
10-2 air

10-3
0.01 0.1 1 10
Photon energy (MeV)

Figure 19 Mass attenuation coefficients for

different materials.

There are other types of attenuation coefficients

besides the mass and linear coefficients. An
example is the mass energy absorption
coefficient, µen/ρ. Scattered photons (X-ray,
Compton and annihilation photons) can travel
some distance from their production site before
donating their energy in the form of secondary
electrons. They may even escape the absorbing
material completely. Secondary electrons
(photo-, Auger-, Compton-, and pair-electrons),
however, expend their energy in the immediate
vicinity of the primary interaction and
contribute to the dose absorbed in this region
(absorbed dose is defined in the next section).
To calculate the dose absorbed by these charged
particles, the mass energy absorption
coefficient, µen/ρ can be used. It is the product
of the mass attenuation coefficient µ/ρ and the
fraction of the photons' energy f, which is
converted to secondary electrons that lose their
energy locally.

µen /ρ = (µ/ρ) f

It is evident by definition that µen/ρ is less than

µ/ρ (µen /ρ < µ/ρ).

29
 CHAPTER 4
Dosimetry
Absorbed dose
Ionization radiation carries energy. In different
processes this energy is transferred to matter.
Some of this energy is deposited locally; some
travels away and might even escape the
irradiated mass volume. Since some ionizing
radiation can participate in nuclear reactions,
energy may also be created (or lost) inside the
irradiated volume.
Ein ∆m Eout
Figure 1 Absorbed energy ∆E in a mass
element ∆m
Generally we may write
∆Ei = Ei,in -Ei,out +Qi (1)
where
Ei,in = energy of an incoming ionizing particle
(excluding rest mass)
Ei,out = sum of energies of all ionizing particles,
created by incoming particle, leaving the place
of interaction (excluding rest mass)
Qi = accounts for changes in the rest mass
energy of the nucleus and of all elementary
particles involved in the interaction.
The energy, ∆E imparted to a volume of matter
with weight ∆m (this volume is regarded as the
place of interaction) is then
E = Σ Ei (2)
and the specific energy imparted, which is also
called absorbed dose, is
D = E/m (3)
30
The absorbed dose D has the unit gray (Gy)
equal to 1 J per kg. An old unit is
1 rad = 0.01 Gy
One may reflect on the size of the unit Gy. A
whole body absorbed dose of 4 Gy to man is a
lethal dose, but raises the body temperature by
only 0.001 oC. Obviously, 1 Gy is a rather large
unit. Local absorbed doses of several Gy are
given in radiation therapy whereas in laboratory
work and in radiation protection, mGy and µGy
are more commonly used.
Relative Biological Effect, RBE
The absorbed dose defined in equation 3 is a
macroscopic unit representing the mean energy
to a rather large mass volume. It does not
consider the stochastic ionization pattern of the
energy delivery. However, low LET and high
LET ionizing radiation can for the same
absorbed dose cause quite different biological
effects due to their different ability to cause
single and double strand breaks. Another
concept, Relative Biological Effectiveness
(RBE) is introduced to describe this situation.
α
Number of chromosome abberations
8
γ
6
4
2
Dα Dγ
0
1 2 3 4
Absorbed dose (Gy)
Figure 2 A comparison of two different
radiation qualities, α and γ. Photon radiation is
always used as the reference since all the
produced electrons are low LET-radiation.
A chosen biological system is irradiated and a
biological effect is measured (in Figure 2 this is
the number of chromosomal aberrations). Two
qualities of radiation are used to irradiate the
system (in Figure 2, α and γ) so that the same
biological effect is obtained. The relative
Biological Effectiveness (RBE) is then defined
as a ratio between the absorbed doses delivered
RBE = Dγ/Dα (4)
Photon irradiation (X-rays > 2-3 MeV or 60Co-
gamma rays) is usually used as a reference. This
is because photons produce a wide range of
electrons in a reproducible way. Electrons of all
energies can be considered as low-LET
radiation. Hence, one should expect RBE-values
close to one for all photon irradiation. This is
also found to be true, maybe except for very low
X-ray irradiation since low energy electrons (<
100 keV) have a somewhat higher stopping
power than higher electron energies (> 100
keV).
Dose equivalent
The RBE-values vary with radiation quality and
chosen biological system. In radiation therapy,
where acute radiation effects are expected, the
RBE value has to be carefully measured for
each situation. The biological effects of
neutrons, protons and heavy ions have to be
compared with established photon and electron
therapy not just in one biological system, but in
many. In radiation protection where radiation
doses are lower and mainly late effects (such as
the risk for induced cancer) are expected, we
may generalize somewhat more.
20
Quality factor, WR
15
10
5
01 10 100
Stopping power in water (keV/µm)
Figure 3 The quality factor WR as a function of
stopping power in water
The measured RBE-values vary from 1 to 20 as
a function of the stopping power of the
radiation. A weighted curve of measured RBE
values (WR) that can be used for radiation
protection purpose is shown in Figure 3.
If we multiply the absorbed dose, D, with WR
we obtain a new dose variable, the dose
equivalent, which includes both the energy
concentration of the radiation and also its
capability to cause late biological effects.
H = WR * D (5)
The unit for the dose equivalent is Sievert (Sv)
if absorbed dose is expressed in Gy.
Effective dose
The radiation sensitivity of different organs in
the body varies. Tissues containing proliferating
cells, such as the bone marrow and the intestine,
are generally more radiation-sensitive than e.g.
the liver. If the radiation dose is unevenly
distributed in the body, we need to consider this
when risks of biological damage are estimated.
For radiation protection purposes, relative
values for different organs are given below
(Table 1). These weight factors mainly relate to
the ability of ionizing radiation to create late
effects such as radiation-induced cancer.
Table 1 Weighting factors recommended by
ICRP 30 for calculation of effective dose.
Organ Weighting
Factor, WT
Gonads 0.25
Breast 0.15
Red marrow 0.12
Lung 0.12
Thyroid 0.03
Bone surface 0.03
Remaining 0.30
1000 We can then define the effective dose as
He = Σ WT * WR * Dorg (5)
where we sum over the different organs. The
unit for the effective dose is also Sievert (Sv) if
the absorbed dose, D, is expressed as Gy. The
calculation of the effective dose is illustrated in
the following example.
31
Example 1. We want to compare the risk of late
effects of two persons exposed to radiation of
different quality and distributions. One is
exposed to 1 Gy of gamma rays, whole body
and the other to 1 Gy of alpha particles by
inhaling radon gas.
Dint (Gy) = 1.6*10-13 *n E β /m (7)
where E β is the mean energy in MeV of the
beta energy emitted per decay, and n is the total
number of decays in the mass m.

Type of γ α Comments For radioactive sources, the absorbed dose rate,

irradiation not the absorbed dose is of interest. Eq 7 can
Absorbed easily be rearranged to
dose (Gy) 1 1
WR 1 20 WR for α is 20 dH/dt = 1.6*10-13 * WR * n/h * E β /m (8)
WT 1 0.12 Ra-gas gives a local
irradiation of lung where

Effective 1 2.4 The Ra-exposed person
dose (Sv) has a 2.4 higher
probability to express
late radiation effect
The effective dose is a theoretical concept used
to calculate risks of various types of irradiation
situations. In practical radiation protection work
one need to define other measurable variables.
Radiation protection instruments are calibrated
in a sphere (tissue equivalent material) of
diameter 30 cm. They are placed 10 cm or 0.07
cm from the surface to give
Hp (10) = individual deep dose equivalent
Hs (0.07) = individual surface dose equivalent
Relation between dose and radioactivity
The energy emitted in radioactive decay can, of
course, be expressed in units of Gy and Sv.
However, since the radioactivity is usually
known in Bq and the energy of emitted particles
in MeV, we need to relate the absorbed dose
from radionuclides in these units.
dH/dt is the dose equivalent rate in Sv/h
WR is the weight factor for radiation quality
n/h is the number of absorbed particles per hour
E β is the mean energy in MeV per particle and
m is the mass unit in kg.
The weight factor WR is 1 for radioactive
decays emitting only beta, electrons and
photons.
A more general treatment of the radioactive
decay is given in the following.
Consider a radioactivity concentration A in an
organ. The radioactive decay emits particles of
different types i = 1, 2, 3 … The mean energy of
particle type i emitted per decay is then
∆i= k ni Ei
where
k is a unit dependent constant
ni is the mean number of particle i per decay
Ei is the mean energy of particle i per decay

The relation between the two energy units Joule ∆i has the dimension energy and can by the
and eV are proper choice of k be expressed in joules (J).
However, usually the dimension is given as
1 eV = 1.6 * 10-19 J (6)
(kg * Gy)/(Bq * h) or (g * rad) / (µCi * h)
From this relation, we can derive a simple
formula for particle irradiation, such as pure The time-integral of the radioactivity
beta-emitting radionuclides concentration in the organ, ∫A(t)dt has the

32
dimension Bq/kg * h. If we multiply the time
integral with ∆i we get the absorbed dose of that
organ if the absorption of the emitted particle is
100 % localized in the organ.
Sources
Figure 4 Distributed radioactivity in the body
gives a rather complicated irradiation pattern.
The radioactivity gives radiation that is locally
absorbed (low energy beta, electrons and X-
rays) and radiation that is partly locally
absorbed (gamma, high-energy beta and
electrons). This radiation may irradiate other
organs (Figure 4) or may completely escape the
body. A complete absorbed dose calculation
after the administration of radioactivity in vivo
also has to consider the kinetics of the labeled
compound and its metabolism and excretion.
If the emitted particle is not locally absorbed we
can give a factor fi (the fraction locally
absorbed) by which we multiply the ∆i -value.
In the body, the situation becomes rather
complicated.
To facilitate dosimetry calculations in nuclear
medicine and in radiation protection, a
dosimetry concept (MIRD) has been developed.
MIRD stands for Medical Internal Radiation
Dose. An expert committee has agreed on
certain standard ways to express and calculate
data. Measured physiological and kinetic data
from human studies are used as input into
theoretical kinetic models, which allows a
33
detailed calculation of integrated radioactivity
concentrations in different organs.
Mathematical standard phantoms have been
derived which allow the use of Monte Carlo
calculation techniques.
Figure 5 A MIRD phantom with standardized
organ sizes and positions. Each organ is given
a mathematical definition that makes it easy to
represent it in a computer. The ∆i-values can be
calculated for each radionuclide using nuclear
spectroscopic data. The fraction of absorbed
energy per organ and the fraction of energy
absorbed in other organs can be calculated as a
function of particle type and energy using
Monte Carlo techniques. From physiological
models, the time integral radioactivity curves
can be calculated for each organ. The dose
equivalent can be calculated for each organ
and the effective dose by summing over all
organs.
Using such techniques, one can rather precisely
estimate the effective dose corresponding to a
given radioactivity of a certain radionuclide.
However, one has to remember that the MIRD
concept still uses a number of approximations,
such as standardized organ geometry and a
homogeneous distribution of the radioactivity in
each organ.
 approximate answers to radiation protection
Table 2 ALI values (Bq) for some common problems and to plan experiments. A number of
radionuclides. The ALI-values correspond to an simple relations have been derived from
effective dose of 50 mSv. equation 8 for different, typical irradiation
situations.
Nuclid ALI Nuclid ALI Nuclide ALI
3
H 3*109 65
Zn 1*107 129
I 2*105 Dose rate on the surface of a beta source
14
C 9*107 69m
Zn 2*108 130
I 1*107
18
F 2*109 67
Ga 3*108 131
I 1*106 The irradiation situation is defined in Figure 6.
22
Na 2*107 68
Ga 6*108 132
I 1*108
24
Na 1*108 73
As 6*107 129
Cs 9*106
32
P 1*107 74
As 3*107 130
Cs 2*109
33
P 1*108 75
Se 2*107 131
Cs 8*108
35
S 8*107 76
Br 1*108 134
Cs 3*106
36
Cl 9*106 77
Br 6*108 134m
Cs 4*109 C Bq/g Dose rate, Hβ/h
38
Cl 6*108 82
Br 1*108 137
Cs 4*106 on the surface
42
K 2*108 81m
Rb 9*109 131
Ba 1*108
43
K 2*108 81
Rb 1*109 133m
Ba 9*107
45
Ca 3*107 86
Rb 2*107 135m
Ba 1*108 Figure 6 A thin-walled tube contains a
47
Ca 3*107 88
Rb 7*108 140
La 2*107 radioactivity concentration of C Bq/kg. What is
51
Cr 7*108 89
Rb 1*109 169
Yb 3*107 the surface dose rate Hβ/h?
52
Mn 3*107 85m
Sr 8*109 192
Ir 8*106
52m
M 1*109 85
Sr 6*107 198
Au 4*107 We use equation 8
54
n Mn 3*107 87m
Sr 1*109 197
Hg 2*108
56
Mn 2*108 89
Sr 5*106 203
Hg 2*107 dH/dt = 1.6*10-13 * WR * n/h * E β /m
52
Fe 3*107 90
Sr 1*105 201
Tl 6*108
55
Fe 7*107 90
Y 2*107 204
Tl 6*107 WR is equal one for beta particles, E β is
59
Fe 1*107 99
Mo 4*107 210
Pb 9*103
56
Co 7*106 99m
Tc 3*109 212
Pb 1*106 about Emax/3 and n/h = C*m*fi.*3600. We then
57
Co 2*107 109
Cd 1*106 210
Po 2*104 obtain
58
Co 3*107 115
Cd 3*107 226
Ra 2*104
60
Co 1*106 111
In 2*108 232
Th 4*101 dH/dt = 1.92*10-10 * C * fi * Emax (9)
63
Ni 6*107 113m
In 2*109 238
U 2*103
64 This is the dose rate if all the particles emitted
Cu 4*108 124
Sb 9*106 241
Am 2*102
67 are absorbed. However, on the surface of the
Cu 2*108 123
I 1*108 244
Cm 4*102
62 source about half of the particles are emitted
Zn 5*107 125
I 1*106 252
Cf 1*103
inwards and do not contribute to the dose
equivalent. Therefore, we obtain
The effective dose has been calculated for many
radionuclides using the MIRD concept. The dH/dt = 10-10 * C * fi * Emax (10)
International Committee on Radiation
Protection (ICRP) has introduced a radiation where
protection variable, ALI (Annual Limit of
Intake) which has the unit of Bq. One ALI is the dH/dt is the dose equivalent in Sv/h
amount of radioactivity that corresponds to an C is the radioactivity concentration in Bq/kg
effective dose of 50 mSv. Emax is the maximum beta energy in MeV
fi is the number of emitted beta per decay that
Approximate dose calculations have the maximum energy of Emax
In the practical laboratory work, simpler
calculation methods are needed to obtain

34
Point source
A common situation is that you work with a
concentrated radioactive source that can be
regarded as a point source. At some distance,
this source gives a dose rate that is related to the
radioactivity amount, and the type and energy of
the emitted particles.
d ∆d
PP
Figure 7 A schematic figure of the radiation
geometry around a point source P, which could
either be a beta source or a gamma source.
The radioactivity is A Bq. The distance from the
point source to the body is d meter. Particles
are not absorbed until they hit the body surface.
A sphere is drawn with the point source at the
center and with the radius d.
Generally, the emitted particles are distributed
on a sphere with the surface S=4πd2. They lose
some of their energy ∆E in a thin layer ∆d with
density ρ. This thin layer has the mass of
4π*d2*∆d*ρ. The number of particles emitted
during one hour is f*A*3600. WR =1 for beta
and gamma. The density of tissue is 1000
kg/m3. If we put these values in equation 8 we
obtain
dH/dt=1.6*10-13 * f*A*3600*∆E/(4π*d2*∆d*ρ)
Rearranging this equation will give
dH/dt=0.458*10-13 *(f*A / d2)*∆E/∆d (11)
35
A beta point source
∆E/∆d is the stopping power of the electrons.
As seen from Figure 8, the stopping power for
high-energy electrons is about constant, about 2
MeV/cm (200 MeV/m). Only for low energies
(< 300 keV) does the stopping power increase.
However, these energies are not interesting
from the external radiation point of view since
their depth penetration is low (< 0.6 mm). Only
the high-energy electrons can penetrate skin and
protection gloves and represent a biological
hazard.
100
300 keV
MeV/cm
10
2 MeV/cm
1
0.01 0.1 1 10
MeV
Figure 8 Stopping power for electrons as a
function of particle energy. Electrons with
energy > 300 keV have about the same stopping
power, 2 MeV/cm.
If we put the value 200 MeV/m of the electron
stopping power into equation (11) we obtain the
approximate relation
dH/dt = 10-11* A*f*/d2 (12)
The dose rate is given as Sv/h, A in Bq and d in
meters.
A gamma point source
If we in Figure 7 change the external irradiation
from beta to gamma, we then have to change the
stopping power for charged particle to
something relevant for photons. A radioactive
decay can also emit gamma rays with different
energies (Ei) and number per decay (fi), which
changes equation (11) accordingly.
The gamma γi with the energy Ei will have
some probability to lose its energy when passing
through the thin layer ∆d. We denote the total
energy that is entering the surface S with Eini
and the energy coming out from the surface
with Eouti. We can then derive the following Table 3 Gamma constants for some important
relation radioactive nuclides, 10-12 Sv*h-1*Bq-1*m-2

Eouti = Eini e-µeni* ∆d (13) Nuclide Γ Nuclide Γ

22Na 0.32 99mTc 0.016
The energy absorption coefficient, µeni, used in
the equation is related to the transfer of energy
24Na 0.47 103Ru 0.090
from photons to charged particles that are 28Al 0.23 110mAg 0.38
locally absorbed. 38Cl 0.19 122Sb 0.064
42K 0.035 124Sb 0.026
Since µeni*∆d is a small value, we can 46Sc 0.29 125I 0.037
approximate equation 13 to
51Cr 0.0043 131I 0.056
Eouti = Eini – Eini*µeni*∆d (14) 54Mn 0.13 134Cs 0.23
56Mn 0.22 137Cs 0.086
59Fe 0.16 140Ba 0.33
The energy released in the tissue volume is then 58Co 0.14 140La 0.30
60Co 0.34 141Ce 0.013
∆Ei = Eini - Eouti = Eini*µeni*∆d (15)
65Ni 0.083 160Tb 0.14
and 64Cu 0.032 177Lu 0.0027
65Zn 0.073 182Ta 0.18
∆Ei/∆d = Eini*µeni (16) 69m-69Zn 0.064 187W 0.080
75Se 0.051 192Ir 0.12
From equation (11) we then obtain
76As 0.064 194Ir 0.040
∆Hi/dt =4.58 10-14 *A*fi*µeni *Ei/d2 82Br 0.039 198Au 0.062
(17) 86Rb 0.013 201Tl 0.012
95Zr 0.11 203Hg 0.035
We can gather the decay-dependent parameters
99Mo 0.038 226Ra 0.22
into a constant
Some considerations about the attenuation
Γ=ΣΓi = 4.58*10-14 * ΣEi * fi* µeni (18)
coefficient
where
The energy absorption coefficient used in the
calculation of the Γ-constant gives the transfer
Ei is the energy of gamma i in MeV
fi is the number of gamma i emitted per decay of energy from photons to charged particles that
are locally absorbed (Figure 9). The following
µeni is the energy absorption coefficient in m-1
example demonstrates how the Γ−constant is
and rewrite equation (17) as calculated.

dH/dt = Γ ∗ A / d2 (19) Example 2. Calculate the Γ−constant for 60Co,

which has two gamma rays, 1.173 and 1.332
MeV. Both gamma rays are close to 100 % per
where
decay. The mass energy absorption coefficient
in water obtained from Figure 9 is 0.03 cm2/g.
the equivalent dose rate dH/dt is in Sv/h,
The density in water is 1 g/cm3 which gives an
the radioactivity A is in Bq, and
energy absorption coefficient of 0.03 cm-1 or 3
the distance d is in meter.
m-1. We use formula (18) to obtain

36
 10000
−12
Γ=4.58*10 -14
* 2.505* 3 = 0.34 ∗ 10 LEAD
1000

The Γ− constant can be calculated for a number

Mass attenuation (cm2/g)

100
of radionuclides as seen in Table 3.

The theoretical background for the attenuation 10

coefficient is given in Chapter 3. The diagram

in figure 9 and figure 10 gives a somewhat 1

simplified information with two curves, which

gives the sum of a number of involved 0,1
processes. Here we will give some examples
how these curves are used in practical 0,01
calculations. 0,001 0,01 0,1 1 10 100
Photon energy (MeV)
For each element, we need to have a special set
of information. Here we will only deal with two Figure 10 Mass attenuation coefficients for
elements, water and lead. photons in lead. Two different attenuation
coefficients are given. The upper curve is the
10000 total mass attenuation and the lower the mass
energy absorption coefficient
1000
WATER
In the calculation of the Γ-constant for 60Co we
Mass attenuation (cm2/g)

100 used the lower curve in figure 9. Water is

usually a good approximation for tissue. We
10
wanted to know the amount of locally deposit
energy and used in this case the mass energy
absorption coefficient.
1

0,1

0,01
0,001 0,01 0,1 1 10 100
Photon energy (MeV)

Figure 9 Mass attenuation coefficients for

photons in water. Two different attenuation
coefficients are given. The upper curve is the
total mass attenuation and the lower the mass
energy absorption coefficient

Figure 11 The degradation of photon energy in

a thick radiation shield.

37
If we would like to calculate the thickness of a 10000

lead container to decrease the radiation from a LEAD

strong 60Co-source 100 or 1000 times, then we 1000

Mass attenuation (cm2/g)

have another situation.
The thick shield can be divided into a number
of thin layers. In each layer we can apply the
mass energy absorption coefficient to calculate
how much photon energy is transferred to the
next layer. However, this energy now consists
of the original gamma energy and of lower
energy Compton photons. The Compton
photons will be, due to their lower energy, more
easily absorbed than the higher original energy
in the next layer. So coming out from the thick
shield will finally mainly be the original gamma
photons, which have undergone no interaction
in the shield and some Compton photons mainly
produced in the last layers.
The energy coming through a thick shield is
then the number of undisturbed high-energy
gamma (can be calculated using the total mass
attenuation coefficient) times the gamma-
energy.
Example 3 A strong 60Co source (100 GBq)
needs to be protected in a lead shield. An
acceptable dose rate from the protected source
would be 2 µSv/h at 1 meter distance. How
thick should the lead wall be?
The dose rate (0.034 Sv/h) of an unprotected
source at 1 meter can be obtained using the Γ-
constant found in table 3. This is
34000/2=17000 times larger than the wanted
dose rate. We need then to reduce the number of
60
Co-gamma by this factor.
100
10
1 E=1,2 MeV
0,1
M=0,06 cm2/g
0,01
0,001 0,01 0,1 1 10 100
Photon energy (MeV)
The two gamma photons from 60Co are rather
similar in energy so we use a value in-between,
1.2 MeV. From the diagram we get (using the
total mass attenuation coefficient) the value
0.06 cm2/g. We multiply with the lead of 11, 3
g/cm3 to get the total linear attenuation
coefficient 0.68 1/cm.
We can then use the exponential relation
between incoming and outgoing photons.
N/No = 1/17000 = e-0.68*X
X is the wanted lead thickness.
X = ln(17000)/0.68 = 14.3 cm
We have made some approximations to obtain
this answer. One is the use of a single energy
instead of doing separate calculations for the
two gamma photons. The other is the use of the
total attenuation coefficient, which gives a
somewhat to small value. The answer could
then be around 15 cm.

38
 CHAPTER 5
THE EFFECT OF RADIATION ON HUMANS
LOSS OF HAIR
3 -4 Gy
DEATH OF BONE
MARROW TISSUE
Renewal of blood cells
stops
3-4 Gy
DEATH OF BOWEL
TISSUE
Renewal of bowel cells
stops
10-15 Gy
STERILITY
1-5 Gy
REEDENING OF SKIN
6 Gy
BLISTERS and
WOUNDS
10 Gy
Acute effects of irradiation in humans
Biological System
Higher organisms are composed of numerous
cells, which co-operate and share the various
functions that they must fulfil. A human being
is formed of some 1014 cells. Some of them, the
nerve cells, are responsible for signal
communication within the organism, while
blood cells and other cells of the circulatory
system transport oxygen and nutrients and some
cells such as kidney cells deal with excretion.
Cells are the smallest units concerned with this
division of labor and for an organism to
function, it is essential for particular cells to be
correctly located and perform their function.
39
Control of the cell is managed by the nucleus of
the cell which contains deoxyribonucleic acid
(DNA), the hereditary material or "memory
store" of the cell. DNA functions as a template
for the formation of ribonucleic acid (RNA),
which in turn acts as the template for the
formation of various protein molecules which
are workers within the cell and perform specific
functions.
Further, there is flow of information from DNA
to RNA to protein. Both DNA and RNA are
carriers of information. DNA keeps information
in the form of a special sequence of nucleotides
– the building blocks of DNA. There are four
different types: adenine, thymine, guanine and
cytosine, designated A, T, G and C respectively.
In a human cell, the total number of nucleotides
in DNA is about five thousand million.
Figure 1 Schematic drawing of a cell and its
most important organelles.
In the double spiral of DNA, A is combined
with T, and G with C. Thus, the information is
found in two arrays, one being complementary
to the other. This can be regarded as "back up"
of the information – something which nature
practiced long before humankind began using it
in our computers. In RNA, nucleotides are
combined in a similar way but with the
difference that uracil (U) is used instead of T.
Three consecutive nucleotides in RNA
correspond to a specific amino acid for forming
protein molecules.
Some examples of different functions of the cell
are described below. Mitochondria are cell
organelles that specialize in converting energy.
They break down energy-rich sugar molecules
and convert the energy into small packets of
adenosine triphosphate (ATP). The different
functions of the body like muscle and cellular
processes are then using ATP for energy
support. Protein synthesis in the cell occurs in
the ribosome with RNA as the template. Storage
of proteins for export occurs in the Golgi
complex. Detoxification of harmful chemicals
takes place in the endoplasmic reticulum and
various substances are broken down in the
lysosomes.
Figure 2 The flow of information in a cell and
comparison with ionization density in sparsely
ionizing (e-) and densely ionizing (α) radiation.
Many other functions are fulfilled within this
small unit with a diameter of 10 – 20 µm.
Different environments in the organelles can be
maintained by membrane separation. Hence,
appropriate conditions exist for simultaneous
energy turnover in the mitochondria, duplication
of DNA in the cell nucleus and break down of
proteins in the lysosomes.
Of the cell content are 60-70 % water and about
15 % protein. The RNA content is a few percent
and less than 1 % is DNA. Large molecules –
macromolecules – which form structures in a
biological system, include DNA, RNA and
proteins. From the radiation point of view,
DNA is the most important biological molecule.
Each cell contains some 5 · 10-12 grams or 1.5
meters of DNA whose diameter is only 2 nm.
40
Energy transfer – primary damage
The effect of radiation on living cells is usually
caused by charged particles like electrons (beta,
Compton or photoelectrons) or alpha particles.
The transfer of energy occurs very quickly –
within pico-seconds. Two types of primary
biological effects can be distinguished: direct
and indirect. In the direct type, the radiation
interacts with biomolecules to cause an initial
modification of their DNA, RNA or proteins. In
the indirect type, which is the more usual in
living cells, the primary changes are in water.
Transfer of energy to the cells' molecules occurs
almost randomly. Hence, it can be expected that
in 60-70 % of all primary events, water receive
the energy. The amount of energy transferred to
the water molecule is indeed small but
significant since it undergoes ionization and
loses an electron. By restructuring, the water
radicals ·OH and ·H, and hydrated electrons are
formed.
Direct action Indirect action
e- e-
Changes in DNA
Figure 3 Transfer of radiation energy to
material in a cell
Radicals are very reactive and react with each
other or with biomolecules within a short time.
If they react with each other they form water,
hydrogen peroxide or hydrogen gas. Water
radicals can react with all biomolecules and
after irradiation, a spectrum of various changed
biomolecules is obtained. The reactions with the
water radicals occur in less than milliseconds.
The energy deposited in a cell is not distributed
uniformly but often in "clusters" (in clumps) of
numerous ionization events and accompanied
by primary damage within a small volume.
When an electron passes a cell, the close dose is
about 3 mGy and the number of primary injuries
to biomolecules is about 100 in the cell nucleus
(the critical organelle of the cell where DNA
occurs). In Sweden, where the background
radiation is 1 mGy (1 mSv) per year, only every
third cell is hit yearly by an electron and the
other cells are unaffected. At very low doses, a
step-by-step increase of radiation dose occurs at
the cellular level, i.e., each cell is hit zero, one,
two or three times etc. The radiation dose to a
larger part of the body, such as an organ, is then
the mean dose for all included cells.
100 ionizations
in the nucleus
High-LET ionizing radiation, such as alpha
particles, creates several ionization events per
nm and therefore very probably creates double
strand breaks. In practice, high-LET ionizing
radiation increases the biological response and
creates more biological damage per physical
dose unit. Permanent double strand breaks can
occur in DNA (fragmentation) or the
information on both strands can be changed in a
small segment of DNA, a mutation.
-17
10 seconds Energy absorption with subsequent
ionization or excitations
-15 -14
10 – 10 Changes in biomolecules caused by direct
or indirect effects
-3
10 to Biological mechanisms of repair splicing
minutes affect the molecular damage
The time scale for following processes is
considerably longer

Information changes Submicroscopic

in the cell injuries
Figure 4 The distribution of energy transferred Hours Microscopically
(ionization events) along an electron track and Mutation visible injuries
when an electron passes through a cell.
Genetic Somatic
damage damage
Cell death
Days
Death of
organisms
Weeks

Years Leukaemia
Cancer

Generations Genetic
damage
to the
offspring

Figure 5 Repair of DNA in a cell and the ability

to repair after different types of irradiation.
As DNA is the critical molecule, the damaging
effect of different types of radiation varies
according to how the energy is distributed in the
molecule. Low-LET radiation creates mainly
single strand damage, which most cells can
repair effectively and accurately. At a low dose,
it is unlikely that such low-LET radiation
creates, simultaneously, two ionization events
within a few nanometers to create a double
strand break.
Figure 6 Time sequence of processes induced in
a cell by radiation.
A repair system exists, which functions more or
less effectively for most injuries to DNA, when
the injury is confined to one strand. This strand
is cut and a small portion broken down and is
then repaired using the other complementary
strand as template whereupon the split in the
DNA is closed and the DNA repaired. Besides
this simple repair system, several more or less
sophisticated systems for repairing damage to
the DNA exist. Since DNA is particularly
important for the cell, many endogenous

41
systems have been developed during evolution
to prevent changes or loss of information (see
also Chapter 6).
Relative Biological Effect RBE
The same dose, irrespective of the radiation
type, produces different effects depending on
the density of ionization. The relative biological
effectiveness (RBE), defined in Chapter 4 is a
way to express this. The RBE can be
determined in many ways and often gives
different values depending on which effect is
studied.
Thus, the RBE for a given type of radiation is
not constant and can, for alpha radiation, vary
from 2 up to 10 - 20. From the different RBE
values that are reported, the weighting factors
for different types of radiation are derived.
Often these are based on some of the highest
RBE values that have been observed. This
weighted RBE is called the quality factor and
usually denoted as WR (Q in older literature).
Acute effects on organisms
Acute effects appear relatively soon after an
irradiation. They can arise only after high doses
of radiation, and in the case of human beings
generally, within two months. This occurred in
the Chernobyl disaster in which 29 people died
because of acute radiation damage. In the 1940s
and 1950s, some people died from radiation
after receiving relatively high radiation doses in
connection with experiments with fissionable
material. A few have died because they were
exposed to radiation from sealed radioactive
sources that had gone astray. In Hiroshima and
Nagasaki, many died from irradiation although
most died from burns or mechanical injury.
Because of the chaotic experiences and
numerous events following the nuclear bomb
explosions, people’s acute response to
irradiation could not be confirmed.
People involved in radiological work and
obeying safety rules are exposed to doses well
below those giving any acute effects. Persons
exposed to radiation doses below 0.5 Gy will
hardly notice it if they do not carry a dosimeter.
42
If a whole body radiation dose exceeds 1 Gy
and is given during a relatively short time (less
than 1 hour) some people will feel sick and
dizzy and have bouts of vomiting. The duration
of these symptoms depends on the radiation
dose. Early symptoms may give some idea of its
magnitude.
Acute radiation damage in multi-cellular
organisms is due to more or less widespread cell
inactivation. Some cell types in the human body
are more prone to cell inactivation than others,
which results in some organs being unable to
function. The law of Bergonie and Tribondeau
often applies; "Cells undergoing division
(proliferating cells) are more radiosensitive than
non-proliferating cells". The organs in which
the formation of new cells is important include
the red bone marrow where blood cells are
formed, the intestine – particularly the small
bone, the spleen and lymph nodes, the skin and
testicles. These organs are also the most
radiosensitive in our bodies. In the red bone
marrow, some 1011 cells are produced daily and
about the same number are produced in the
intestine. After intense radiation with some Gy,
new formation of cells in the bone marrow will
be reduced drastically and furthermore many
bone marrow cells will die through apoptosis.
The same is true for the small intestine,
although at higher doses.
The cause of death in people exposed to whole
body irradiation for a short time with doses
between 3 and 10 Gy is found in the bone
marrow. These doses inhibit the development of
cells and cause marked reduction in the number
of cells, which have a short life span.
Thrombocytes
Granulocytes and lymphocytes
Stem
cells
Red blood cells
Figure 7 Blood cells develop in the red bone
marrow. The number of granulocytes and
thrombocytes in the blood are influenced
markedly by high doses of radiation.

The life span of red blood cells is about 120
days, which means that their reduction in
number is small. White blood cells such as
granulocytes and lymphocytes have a much
shorter life span (some days). Hence, any arrest
of their formation very quickly results in
marked reduction in their number in circulating
blood. Lymphocytes are highly radiosensitive
and become markedly reduced after irradiation,
partly due to reduction of stem cells in the red
bone marrow but also due to extensive
apoptosis. These cells are important for the
immuno-defence of the body and irradiated
individuals become therefore very infection
prone.
Thrombocytes are important for the coagulation
of blood. Small ulcers and considerable
hemorrhages arise in the mouth, intestine and
skin. The number of thrombocytes is also
markedly and quickly reduced after irradiation.
People exposed to whole body radiation doses
of 3 to 5 Gy may die within 3 - 4 weeks. About
50 % of those irradiated with 4 Gy survive.
Bone marrow transplantation may improve the
prognosis after exposure to doses of between 3 -
10 Gy and was carried out in a number of
people irradiated in the Chernobyl disaster, but
with limited success. Bone marrow
transplantation necessitates that an
immunologically compatible bone marrow
donor is available. In some hospitals, bone
marrow transplants are used routinely after
radiation therapy for the treatment of certain
leukemia. Purified stem cells from the patient
are then often used.
43
Figure 8 Reduction in the number of
granulocytes and the sequel in humans after
high doses of radiation.
The small intestine is the critical organ at doses
of 10-15 Gy or more. Cells covering the inner
part of intestine continuously migrate upwards
on the intestinal villi. If the producing stem cells
in the crypts are irradiated the development of
these cells protecting cells stops. The cells
already formed continue to migrate but are not
replaced and increasing ulcers develop in the
small intestine. This leads to massive loss of
water and salts, and to an invasion of the
intestine content.
Figure 9 Transverse section of the small
intestine and details of the villi and crypts.
Other acute effects also are generally related to
the influence of proliferating cells, but there are
some exceptions such as the reaction of
reddening and feeling of warmth of the skin,
which depends on increased blood flow.
Effects on the fetus
The dose of radiation required to cause acute
effects in humans is apparently high. Since
proliferating cells are radiosensitive, the embryo
and fetus are particularly sensitive to radiation.
Fetal development is divided into three phases.
The pre-implantation phase extends over some
10 days after fertilization. The growing embryo
is particularly radiosensitive during the early
cell divisions where the ‘all or nothing’ law
applies. This means that if the embryo is
damaged by radiation in this phase, spontaneous
abortion occurs at an early stage but the risk of
malformations during is relatively small.
The organogenesis phase embraces the period
from 10 to 60 days of fetal development. During
this phase the foundation of all the organs
occurs except certain parts of the central
nervous system. Ionizing radiation can cause
fetal injury mainly during a short period when
the organ begins to form and there is active cell
proliferation. The period of high radiosensitivity
for most organs is short. Skeletal malformations
are relatively common after radiation (compared
with malformations in other organs). A slight
increase in the frequency of defects in the
skeleton has been detected after very low doses
(in mice irradiated with 0.05 Gy at day 7).
During the growth period, radiosensitivity is
lower than in earlier stages with one important
exception that concerns the dangerous influence
radiation has on the development of the central
nervous system. This occurs late in fetal life.
Development of the cerebrum is a very
protracted and radiosensitive period compared
with the development of other organs. Cerebral
effect has been confirmed amongst children
born after the explosion of the atom bombs in
Hiroshima and Nagasaki. In an investigation of
1,599 children irradiated during fetal
development, excess in the frequency of mental
retardation was observed.
Figure 10 Frequency of microcephaly, reduced
size of head, in children born after explosion of
the atom bombs in Hiroshima and Nagasaki.
Irradiation occurred during weeks 6 to 11 of
fetal development.
The individuals had difficulty in doing simple
calculations, making simple conversation or
taking care of themselves. Most of these
44
individuals were referred to institutional care.
The risk of serious damage was found to be
greatest if the radiation occurred during weeks 8
- 15 of fetal development.
The result does not exclude the absence of a
threshold value; that is to say, there might be a
linear dose-effect relation. However, it is
important to realize that in the group with low
radiation doses, the number of mentally retarded
was only three and the statistical errors are
large.
The risk of mental retardation, when the fetus
was irradiated in weeks 8-15 of the pregnancy
was calculated to be 0.04 per Gy. This shall be
interpreted as follows: If 1000 fetuses at this
developmental stage are exposed to 1 Gy each,
then 40 of the children will be mentally
retarded. If irradiated after week 16, the risk is
four times less. With low doses of radiation, the
effect on the central nervous system seems to be
the greatest risk.
Figure 11 Frequency of mental retardation in
children irradiated during weeks 8 to 15 of fetal
development.
In Hiroshima and Nagasaki, a smaller head has
been observed to occur with the same increased
frequency as mental disorders. The critical
induction period for the head size occurs earlier
in fetal development and there does not appear
to be a clear association between these two
effects.

Figure 12 The development of cells in the
human cerebrum.
Effects on the development of the central
nervous system after low radiation doses can be
seen in low frequency in experimental animals.
The effects studied included the size of the
cerebrum, the incidence of cell death and
disorientation of nerve cells. Most people
working in this experimental field think that
there is a threshold value somewhere between
0.05 and 0.1 Gy.
Late effects on organisms
Fairly soon after the discovery of X-rays by
Roentgen in 1895 and natural radioactivity by
Becquerel in 1896, it was established that
radiation had harmful effects. Pioneers in the
field of radiation worked without protection and
received very high doses, particularly to the
hands and arms. After reddening of the skin and
slow healing ulcers in the acute stages of the
injury, forms of cancer appeared sufficiently
often for the risk to be noticed.
Genetic effects became evident somewhat later.
In 1927 Muller demonstrated that X-rays could
produce mutations in the banana fly. Other late
effects include opacity of the optic lens and
fibrosis, e.g. collections of fibrous tissue.
Induction of cancer
Quite soon it became apparent that ionizing
radiation could both cause and cure cancer.
45
Early findings indicated a threshold under
which radiation was not harmful and would not
cause cancer. However, with time people
became more and more concerned about the
risks involved. The increasing use of ionizing
radiation in society (X-ray diagnostics and
therapy) also promoted the interest to quantify
these risks.
It is still not strictly proved that the induction of
cancer is without a threshold value. However, it
must be stated that our knowledge of risks with
ionizing radiation is far more accurate than with
any other type of biological hazard. The risk
estimates in radiation protection are
conservative and assume that all doses can
cause biological damage but that the risk is
proportional with the dose – if the dose is
reduced ten times, the risk is also reduced ten
times.
The risk assessment is based on experience of
groups of people who have been irradiated.
1. Survivors of the nuclear bomb explosions in
Hiroshima and Nagasaki in Japan. Knowing
where people were located at the time of the
explosion, the radiation dose has
subsequently been calculated for the
individuals. Of course, individual dose
estimations are associated with gross errors
but are still precise enough to permit risk
assessment.
2. People who had radiation therapy for
relatively benign conditions such as back
and joint complaints. The doses were
correctly determined and entered in their
clinical records along with the region of the
body, which was irradiated (the radiation
field).
3. Miners who were exposed to radon-222 and
its radioactive daughter products. The lungs
are the critical organs in these cases.
4. Children who during fetal development
were exposed to X-rays in connection with
diagnostic X-ray examination of the
maternal pelvic region.

Figure 13 Excess frequency of cancer in
Hiroshima and Nagasaki after the nuclear
bomb explosions.
In addition, several epidemiological
investigations in this field exist. Some of these
comprise a great number of people and
therefore provide important information.
Without doubt, the most important group are the
survivors of the Hiroshima and Nagasaki
nuclear bomb explosions. In investigations of
small groups, results are often obtained that
sometimes suggest increased risk and
sometimes lesser risk from low doses depending
on the random frequency of cancer cases. It is in
such instances important not to draw prejudiced
conclusions from isolated investigations.
Instead evaluations must be made from all the
epidemiological investigations taken together.
In Hiroshima and Nagasaki, at the end of the
40s and beginning of the 50s, an increasing
frequency of leukemia was confirmed.
Later, a number of other types of cancer showed
increased frequency. The total number of cancer
cases related to irradiation was ”only” about
500.
A problem, which is discussed in many
contexts, is the shape of the dose-effect curve at
low radiation doses. Different interpretations
are given but the truth is that it cannot be shown
how the curve looks at low doses. ICPR
(International Commission on Radiological
Protection) states in its risk assessment (low
radiation doses and dose rates) a 5 % risk per Sv
of getting radiation related cancer if a
population of normal age distribution is
irradiated. This implies that if an individual is
exposed to 1 mSv, there is a risk of 1 in 20,000
46
of dying of radiation-induced cancer. For adults
the risk is 4 % per Sv.
During the fetal development the risk appears to
be still somewhat higher. This assessment is
based on examination of children who during
fetal life were exposed to radiation during X-ray
investigation of the maternal pelvic region.
Figure 14 Relation between radiation dose and
excess frequency of leukemia.
Genetic effects
Changes in the genetic information (mutations)
are first apparent in the future generations. The
cause is a change in the information material
(DNA) in the sex cells (spermatozoa and egg
cells). If the change is small, it is called point
mutation and if large, it is usually called
chromosome change.
Quantification of genetic risks is more difficult
than for cancer. For example, an increase in
hereditary damage has not been discerned in the
offspring of irradiated people in Hiroshima and
Nagasaki. This does not mean that the
irradiation did not induce any genetic changes
amongst these people but that no statistical
increase was demonstrated. It is thought that the
risk of genetic injury is less than the risk of
cancer.
Internal irradiation
The risk of external radiation is low when
working with low energy beta-emitting
radionuclides like 3H, 14C and 35S. In fact, these
beta particles can not even penetrate the skin.
However, there is always a risk of internal
contamination by inhaling, through the mouth
or by absorption of lipophilic labeled
compounds.
Generally, it is difficult to make assessments of
the risks because possible uptake and effects
depend on which compound is labeled. The
biological half-life of e.g. 3H depends not only
on its decay but also on the metabolism and
biological half-life of the labeled compound.
Thymidine, labeled either with 3H or 14C, is
incorporated in the DNA of proliferating cells.
Naturally, if 3H is a part of DNA, the critical
biomolecule, the dose effect should be greater
than if it is incorporated into e.g. a protein. Beta
particles from 3H decay have a maximum range
of 7 µm, which is about the diameter of the
nuclei in our cells. One might guess that in this
situation the biological half-life should be long
and thus the radiation risk large. However,
thymidine is rapidly metabolized in the body
and only a few labeled thymidine molecules are
actually incorporated into DNA. Rapidly
proliferating cells incorporating thymidine also
renew and replace their DNA fast. Cells in the
small intestine, for example, are renewed within
a few days. The time the 3H will stay in the
DNA is therefore short.
The risks are greater if the radionuclides are
incorporated into structures that are stable. This
can apply to the incorporation of 3H-thymidine
in the cells of growing individuals in whom the
biological half-life can be very long. The same
is true for radionuclides such as the isotopes of
Ca, Sr, Ba and Ra that are incorporated into
bone tissue. Often, these elements are reused
and are not metabolized in the same way as
organic compounds. Iodine isotopes are mainly
taken up by the thyroid gland and are
incorporated into the thyroid hormone, which is
stored in the gland for a long period and can
give high doses locally.
As a rule only marginal measures can be taken
to reduce the effect of radionuclide intake. The
uptake of radioactive iodine in the thyroid gland
can be reduced by the intake of non-radioactive
iodine but this should, preferably, occur before
that of radioactive iodine. Similarly, the uptake
of Sr-isotopes is reduced if stable Ca is
supplied. In principle, the same thing also
47
happens if inactive thymidine is given together
with 3H-thymidine. But this way to protect
against internal contamination is usually not
very effective and not very practical.
Comparison: Radiation – Chemicals
In certain situations, it is desirable to be able to
compare the risk assessments of radiation and
chemicals that affect us. With the establishment
of limited concentrations of chemicals in our
environment, it is taken for granted that there is
a threshold value. No one can affirm with
certainty whether this threshold value really
exists or whether in most cases, it is tenable. In
general, there is an attempt to stop the use of
carcinogenic substances.
Regarding radiation, since a limiting value is
established it is equivalent to accepting the risk
of cancer but at a very low level. Note that the
natural background radiation gives the same
risk as a similar amount of added radiation
dose. Knowledge of the effects of radiation is as
a rule more widespread than that of chemicals
even if it is usually easier to understand the
effects of chemicals. Radiation hits very non-
specifically, whereas a chemical substance often
has a specific effect. Radiation, however, has
from the beginning been considered dangerous
in that some of the early pioneers developed
cancer and this risk has been documented
further after the Hiroshima and Nagasaki
nuclear bomb explosions. Besides, there are
only a few types of radiation to consider, which
makes epidemiological research easier.
On the question of chemicals, people as a rule
have had a basically different approach – it is
only in the last 10 years that a greater awareness
of their dangers has been revealed. In addition,
there are numerous chemicals and it is
impossible for anyone to have a detailed
knowledge of the dangers of all of them. This is
valid especially when a long-term view is taken.
In most cases, from a risk point of view, direct
comparisons of radiation and chemicals are very
difficult to make. Naturally, there are gaps in
our knowledge of radiation but the gaps are
even greater for chemicals. If a comparison is
made it must proceed from a number of
premises that furnish the final comparison with
large errors.
Radionuclides in nature
The cosmic radiation from outer space consists
of particularly high-energy particles, especially
protons. When cosmic radiation hits the earth's
atmosphere, large numbers of secondary
particles are formed. This radiation can also
create radionuclides such as 14C in the
atmosphere.
A number of radionuclides have such long half-
lives that they have survived since the ”creation
of elements”. Some of these decay to other
radioactive elements. In certain cases, long
chains of decays occur by which a radioactive
substance - by successive radioactive decays - is
converted into one daughter nuclide after
another, until a stable (non-radioactive) end
product is formed. Most radioactive material
occurring in nature is included is one of four
such chains, all of whose links have atomic
numbers over 81.
In Sweden, the largest contribution to natural
background radiation comes from radon-222
and its decay products ("radon-daughters").
They form part of a chain of decay, which
begins with uranium-238 and ends in lead-206,
which is stable.
Besides these chains, some other radionuclides
occur in nature. One example is potassium-40
which forms 0.01 % of all natural potassium.
Another is carbon-14, known for its use in
determining the age of organic archaeological
findings.
The Chernobyl accident
The Chernobyl disaster in 1986 resulted in the
deposition of radiocesium, mainly 137Cs, in the
ground, vegetation and in the lakes in large
areas of Europe. In the worst affected regions
problems arose concerning the production of
food of acceptable quality. Apart from this, the
fall out caused only minor disturbances. SSI
(Swedish Radiation Protection Institute) issued
48
instructions on how people in Sweden should
work with wood ashes and sludge, which in
certain regions was contaminated by high
concentrations of radiocesium.
During the spring and summer of 1986,
comprehensive restrictions and measures were
adopted concerning the agricultural production
of food. The reason for introducing restrictions
was that radiocesium was deposited directly on
grasslands. From 1987 onwards, the uptake of
cesium by their roots caused the contamination
of the crops, which normally resulted in low
cesium concentrations in agricultural products.
In the future, the problem will instead be with
foodstuffs produced in the natural ecosystem.
The radiocesium content in these products now
is much greater than in products of the
agricultural system and besides, diminution of
the content occurs slowly or not at all. The latter
applies particularly to the radiocesium content
in elk and roe deer, which was the same in 1993
as in 1986. Fish in the forest lakes have shown a
slow reduction but in reindeer, the reduction has
occurred more quickly.
The recommendation that is usually given, is
that the radiation dose arising from the intake of
radiocesium from Chernobyl should fall below
1 mSv per year, which corresponds to the intake
of 80,000 Bq of 137Cs. The fallout from
Chernobyl also contained 134Cs and the intake
of 60,000 Bq of this cesium isotope corresponds
to 1 mSv. 134Cs can be disregarded since it
"only" has a half-life of 2 years compared with
30 years for 137Cs. As mentioned earlier, the
risk of 1 mSv is equivalent to 1 in 20,000 dying
sometime in the future from radiation-related
cancer. This is considered to be an acceptable
risk. Note that of these 20,000 persons, more
than 6000 will obtain cancer due to other
reasons.
Dose limits for radiological workers

In different countries legal rules for radiological

workers may vary somewhat. In Sweden, the
authority is SSI (Swedish Radiation Protection
Institute), has given the following limits.

Period of time
Quantity
Limits
(mSv)/year
Workers in Annual
General Effective dose 50
Dose equivalent to 150
the lens of the eye
Dose equivalent to 500
the skin
Dose equivalent to 500
hands, forearms feet
and ankles
In addition, for 5 100
consecutive years.
Effective dose

Students and Effective dose 6

Trainees aged Dose equivalent to 50
16 - 18 years the lens of the eye
Dose equivalent to 150
the skin
Dose equivalent to 150
hands, forearms
Feet and ankles

Table 1 Dose limits for people working with

ionizing radiation from external sources

49
 CHAPTER 6
Radionuclide targeting
where Emean is the mean energy (MeV) of the
INTRODUCTION beta particle (Emean= Emax/3), m is the mass
of the tumor (kg) and N is the number of
Tumor cell-specific targeting for delivery of radioactive decays given by the relation
toxic agents is an attractive approach for the
killing of spread cells whose positions cannot N=A x T½eff / ln(2)
be determined through available diagnostic
methods. The targeting process might not in where
itself give a satisfactory treatment of large A is the radioactive uptake (Bq) and
tumors because the radiation dose may not be (T½eff)-1 = (T½phys)-1 + (T½biol)-1 in seconds.
sufficiently high or the targeting substance may
not penetrate well. However, large tumors with
The formula predicts, for example, that if a total
rather well defined margins can often be treated
of 10 MBq of the radionuclide 90Y is taken up
by surgery, or radiotherapy with external or
in a tumor with the mass 20 g (20 cm3) then the
internal radiation sources. In the latter case,
tumor dose will be about 10 Gy.
encapsulated radiation sources are positioned in
the tissue interstitium or in body cavities.
The formula has to be corrected if a certain
Therapies based on targeting such as radio-
fraction of the electrons escape from the tumor
immunotherapy and boron neutron capture
mass. The same formula applies for alpha
therapy can be complementary with the major
particles if Emax is used instead of Emean and can,
aim to kill spread cells.
in principle, be used also to calculate the dose to
different organs due to background irradiation
The radiation biology considered in connection
by photons. Calculations of photon-mediated
with the use of targeted radionuclides is mainly
doses require that a reasonable assumption can
concerned with the effects of the radiation from
be made about the radioactivity uptake in a
the radionuclides on the targeted cells or their
defined mass of tissue and the average
close neighbors. This means that the targeting
absorption of the photons. It is actually possible
process is mainly based on the desire to
from modern scintigraphy to make reasonable
optimize the biological action of electrons from
assumptions about the relative photon release
beta decays (beta minus and beta plus) and on
from different parts of the body and by knowing
alpha particles from the decay of heavy nuclei.
the total amount of injected radioactivity, it
Photons (gamma- and X-rays), which are used
might be possible to make good approximations
in medical diagnosis and which most often also
of the number of Bq in different body regions.
accompany alpha and beta decays of
radionuclides used for therapy, are not of direct
PRIMARY INTERACTIONS
interest. Photons do not deliver their energy in
the local environment around the targeted cells.
Most of the radiation damage is mediated via
Instead, they deliver their energy to large
ionization of cellular water, which gives free
volumes of the body and give an increased
radicals with the capacity to damage critical
background dose, which is undesirable.
biomolecules. Only a small part of the
radiation-induced damage is obtained through
ABOSRBED DOSE direct ionization of the biomolecules (Fig. 1).
Mainly high-energy beta (e.g. from 131I or 90Y)
DNA is the most important biomolecule when
and alpha particles (e.g. from 211At) are
the action of radiation is considered. The
considered to contribute to the local dose in
formation of radicals is increased in the
targeted radiotherapy. The tumor dose for beta
presence of oxygen, and oxygen can also react
particles can be calculated approximately by
with primary lesions in DNA causing fixation of
damage. Oxygen is in fact a powerful radiation
Dose (Gy) =1.610-13 N Emean / m
sensitizer. Low molecular weight thiols (e.g. SH
containing amines or amino acids) are

50
protecting substances because of their normally
high proton-donating capacity. The main
intracellular protector of this type is the tri-
peptide glutathione
Figure 1 Schematic drawing of the primary
interactions taking place after irradiation. The
sensitizer O2 and the protectors of SH-type act
both at the primary radical formation level and
at the fixation of DNA damage level.
. One Gy of low-LET radiation gives about 1000
a b
c d
Figure 2 Schematic descriptions of proposed
DNA repair mechanisms.
a. Single strand breaks. Taken care of by
excision repair. Exo- and endonucleases
remove sugar-phosphate and bases from the
damage. Polymerases synthesize a new
strand using the intact stand as a template.
Finally, ligase closes the opened ends.
b. Base damage. Glycosylases remove the
damaged bases, which introduces a single
strand break. This is repaired by the
mechanism described above (see a).
c. Double strand breaks. In one proposed fast
mechanism, enzymes trim the two near ends
so that they fit together. Thereafter, the ends
are brought together, similarly to how it is
made in a plasmid using DNA technology.
51
Since parts of DNA are lost, this repair type
is most likely when introns are damaged. In
a slower type of repair, a near homologous
DNA sequence is “lined up”. Strand
exchanges convert the double strand break
into two single strand breaks (one on each
strand) that are repaired by excision repair.
Gene rearrangements or translocations can
occur if the damage DNA sequence
exchanges genetic material in the lining up
process. This process is probably necessary
when exons are damaged.
d. Cross-links. The two DNA strands are
covalently linked. This is a serious damage
since it prevents strand separation at
transcription and DNA synthesis. This
repair mechanism is not well understood,
but probably involves mechanisms similar
to those described for double strand breaks.
DNA DAMAGE AND REPAIR
DNA is the critical target for radiation damage
as has been shown in numerous experiments.
single strand breaks, 1000 base damages, 30-50
double strand breaks and a few cross-links in
DNA. The single strand breaks and the base
damage can effectively be repaired by the
excision repair system as indicated in Fig 2.
Double strand breaks can also be repaired
although the mechanism is not clear and the
(fidelity correctness?) of the repair is not
necessarily high. Errors in the repair of double
strand breaks in DNA might give rise to
rearrangements (translocations) of genes. It is
assumed that, several hours after the irradiation
with a dose of 1 Gy, there are on average one or
two unrepaired or seriously miss-repaired
double strand breaks or cross-links in DNA.
This remaining damage probably gives rise to
inactivation of cell proliferation. If such damage
is randomly distributed among cells and all of
them, with a high probability, inactivate cell
proliferation then the shape of the cell survival
curves in Figures 4-8 can be qualitatively
understood. The only assumption that has to be
made is that the lethal damage is Poisson
distributed among the cells. This gives a certain
probability of survival for each cell independent
of how high the dose is.

INTRINSIC RADIOSENSITIVITY
Knowledge of the factors that determine
intrinsic radiosensitivity is limited. However, it
is known that cells react strongly after radiation
damage. More than 40 genes have been reported
to be activated by ionizing radiation. The
molecular genetic knowledge about the
regulation and trigging of DNA repair is not
known in detail although some insight has been
gained the last years. Figure 3 is highly
speculative but includes some of the most
recent theories of the relations between the
genes involved. The DNA is checked regarding
structural damages by unknown mechanisms
(DCM). When severe such damage is found,
certain genes code for transcript factors. These
activate genes that code for inhibitors to cyclins
and the corresponding cyclin-kinases. This
gives a block in the cell cycle and the cell will
then have a reasonable time for enzymatic
repair of critical DNA damages before it is
forced into mitosis. The DNA-histone complex
is, if necessary, opened in preparation for the
repair enzymes to reach the damaged sites.
Radiation
Cell division
DNA check
mechanism
No! DNA repair
DNA damage ?
DNA-histone
Yes!
opening
Gene activation mechanisms
Transcript factors Growth delay
Gene activation Cell cycle inhibitors
Figure 3 A schematic overview of assumed
mechanisms involved in cellular and molecular
reaction on radiation induced DNA damage.
Some diseases are known to be due to defects in
the DNA repair system or closely related
systems. Ataxia telangectasia is a disease where
52
it is assumed that transcript factors are mutated
or defective, which prevents the slowing down
of the cell cycle. The cells do therefore not have
enough time to repair DNA damage before they
are forced into mitosis. In Blooms syndrome
there is a defect in the ligase activity, and in
Fanconi´s anemia it seems to be difficult to
repair certain DNA cross-links.
The most dramatic increased radiosensitivity is
seen in Ataxia telangiectasia. This indicates that
cell cycle block and cell cycle check points are
critical, not only for genetic stability as
previously has been proposed, but also for the
determination of radiosensitivity. However,
more basic research is needed to obtain better
knowledge about the factors that determine
intrinsic radiosensitivity.
SURVIVAL CURVES
The initial events at the radiochemical level
give rise to different types of biological effects
that can be measured at the cellular level. The
most important effect, from the therapeutic
point of view, is of course the inactivation of
tumor cell proliferation and this will mainly be
discussed below. The arrest of cell proliferation
is mediated through severe damage in DNA.
The shape of cell survival curves is well known
from experiments with external radiation. Cell
survival is defined as the relative number of
cells that, after irradiation, has the capacity to
form colonies containing at least 50 cells. This
is analyzed through the sparse seeding of the
analyzed cells as monolayers or as suspension
cultures in agarose gel. One or two weeks after
irradiation, the exact time being dependent on
the growth rate of the cells, the number of
colonies containing at least 50 cells is scored.
Cell survival is "fractional" in that a certain
fraction of cells always seem to have survived
as shown in Fig. 4. This is due to heterogeneity
in local energy deposition and heterogeneity in
structural DNA damage and repair. Thus, with
an increasing dose the probability of lethal hits
increases for each cell, giving rise to the type of
"fractional" curves seen in Fig. 4.
 Figure 5 Schematic drawings of cell survival
curves with the functional parameters Do, n,
α, β and S2Gy indicated in the curves.

Table 1. The table shows crude estimates of the

Figure 4 Typical clonogenic cell survival functional parameters Do, n, α, β and S2Gy.
curves after irradiation with low- or high-LET Typically, glioma, melanoma and osteosarcoma
radiation. cells are radioresistant (Low), adenocarcinoma
cells are intermediate sensitive (Medium)
MODELS whereas lymphoma, myeloma, leukemia and
Cell survival can be mathematically modelled. small-cell lung cancer cells can be regarded as
Commonly used models are the single-hit, radiosensitive cells (High).
multi-target and linear-quadratic models. The Degree of radiosensitivity
former is described by
Low Medium High
Do (Gy) 1.0 - 3.0 1.0 – 2.0 0.5 – 1.2
S = 1-(1-exp{-D/Do})n
n 2 – 10 2-3 <2
where D is dose (Gy) and Do is a constant (Gy) α (1/Gy) 0.01-0.5 0.03-1.0 0.05-2.0
giving the slope of the curve in the high dose β (1/Gy2) 0.001-0.05 0.001-0.1 0.005-0.2
region. The extrapolation number n gives the
radiosensitivity in the low dose "shoulder" S2Gy > 0.5 0.3 – 0.5 < 0.3
region (Fig. 5a). The linear-quadratic model is
described by the simple relation DOSE RATE

S = exp{-(αD + βD2)} One main difference between external

irradiation and irradiation by targeted
radionuclides is that the latter most often gives a
where α is the slope of the curve near the dose
much lower dose rate. A low dose rate means
zero and β is the constant describing the shape
that critical radiation damage in DNA can be
of the curve at high doses (Fig. 5b). Both
repaired during the irradiation, which gives a
formulas are used to give a simple two-
less efficient inactivation of cell proliferation as
parameter description of the radiosensitivity of
shown in Fig. 6. The shifts in Figure 6
normal and tumor cells.
illustrates the changes in survival seen when the
dose rate is shifted from about 1 Gy/min to
It has been claimed that the survival of cultured
about 1 Gy/hour and further to 1 Gy/day. One
tumor cells at 2 Gy correlates, to some degree,
Gy/hour allows DNA repair to take place during
with the clinically observed radiosensitivity of
the irradiation, which means that more dose is
the corresponding types of tumors. Typical
needed to inactivate the cells.
values of Do, n, α and β are given in Table 1
along with typical survival levels at 2 Gy, S2Gy.

53
 Figure 6
Examples of
cell survival
curves
when the
dose rate is
changed
One Gy/day allows both DNA repair and cell
proliferation to take place during the irradiation
and the latter means that the number of cells
increases during the irradiation, making it
necessary to kill more cells. It is likely that the
dose rate in targeted radiotherapy is so low that
DNA repair, and sometimes cell re-population,
during irradiation have to be taken into account.
The low dose rate makes targeted radiotherapy
inefficient when cell inactivation per dose unit
is considered. However, this does not at all
disqualify targeting of radionuclides for therapy.
The tumor cell specificity is the main critical
aspect and if the tumor cells obtain a low dose
rate, in spite of a good tumor specificity of the
targeting agent, then the normal tissues must
have an even lower dose rate. Thus, normal
tissues will in such cases always have better
possibilities to repair unwanted damage. It is the
differential uptake between normal and tumor
tissue that is important. If the tumor specificity
is too low then it will be difficult to obtain high
tumor doses without also giving too high doses
to normal tissues, independent of the dose rate.
HIGH-LET
Conventional radiation with low ionization
density, low-LET (LET = Linear Energy
Transfer), delivers an ionization density in the
range of 1 eV per nm, which seldom gives
dense clusters of ionization. The corresponding
values for high-LET radiation (e.g. alpha
particles) are 100-200 eV per nm. A delivered
energy of about 30 eV is needed for each
ionization, since about 10-15 eV is needed for
the release of a bound electron and the rest of
the energy is converted to thermal energy.
54
ion
electron
Figure 7 A crude comparison between the
ionization densities and the size of DNA when
low- and high-LET radiation types are applied.
Considering that the diameter of the double
stranded DNA molecule is 2 nm and that high-
LET delivers in the range 200-400 eV over that
distance, it is easy to realize that densely
ionizing radiation can give severe and clustered
damage, which is difficult to repair. High-LET
radiation can be considered to give few particle
tracks, but everything that is traversed is
destroyed. Low-LET radiation gives more
randomly scattered ionization separated by
rather large distances along the DNA molecule
(Fig. 7) and the cell has a good chance to repair
such damage.
There are nearly no dose rate effects when high-
LET radiation (e.g. alpha particles) is applied in
targeted radiotherapy. It does not matter if two
severe damages on DNA are made within a few
hours or within only a few minutes. The damage
can probably not be effectively repaired and
each damage might be severe enough to inhibit
further cell divisions. Thus, provided the uptake
of the radionuclides is tumor specific, it might
be an advantage to use high-LET radiation
because the probably low dose rate is not an
obvious disadvantage. It might in fact be
enough to administer the radionuclides over a
very long time.

OXYGEN
Acute hypoxia or anoxia in a tumor gives
radioresistance due to the decreased number of
formed free radicals and the lack of oxygen-
mediated fixation of damage. When oxygen is
depleted there are better chances for proton
donation from glutathione, which also means
increased protection. The quantitative
determination of the oxygen effect is given as a
dose-quotient as shown in Fig. 8. Chronic
hypoxia can under certain conditions also give
radioresistance.
The problems with hypoxia and anoxia are the
same when targeting of radionuclides and
external radiation are applied. However, one
specific problem relating to targeting is that
hypoxia and anoxia is an indication of bad
vascularization, which might in itself prevent
the targeting agent from reaching all tumor
cells. Thus, it is possible that hypoxia and bad
uptake of radionuclides might correlate. This
has to be analyzed further. Something that is
well known is that high-LET radiation is
equally effective on hypoxic cells and normoxic
cells, which means that it could, in some cases,
be an advantage to use such radiation.
Figure 8 The influence of hypoxia on cell
survival. The method to calculate the oxygen
enhancement ratio, OER, is indicated.
However, it is far from clear to what extent
human tumors suffer from such severe hypoxia
55
that the cells really are radioprotected. The
partial oxygen tension has to be much below 10
mm Hg for the cells to be fully radioprotected.
Another unsolved question is whether severely
hypoxic cells still have the capacity to
proliferate. Furthermore, some scientists have
claimed that even if there are hypoxic areas with
clonogenic cells in certain regions of a tumor,
these cells might not exert a problem because of
reoxygenation during the course of fractionated
radiotherapy. It can be concluded that in spite of
several years of research there are still
uncertainties regarding the importance of
hypoxia for the curability of tumors and the
uncertainties regarding this problem are not
smaller when targeted radiotherapy is
considered.
CYCLING CELLS
.
Figure 9 Examples of reported variations in the
radiosensitivity for resting cells and cells in the
cycle when exposed to low- and high-LET
radiation.
Cells in the cycle are, on average, more
sensitive to low-LET radiation than cells in
resting phase and this applies equally to both
normal and tumor cells. Furthermore, the
radiosensitivity has been reported to vary over
the cell cycle as shown in Fig. 9 when
conventional low-LET radiation is applied and
there is no difference whether the radiation
comes from an external or an internal radiation
source.
High-LET radiation prevents the cells in the late
S and early G1 phases from being protected. In Table 2. Examples of radionuclides of
fact, it is assumed that cells out of the cycle are therapeutical interest. The last column indicates
as sensitive as cells in the cycle when high-LET the labeling mode. Ch stands for chelator
is applied, and it might therefore be an mediated whereas H stands for halogen
advantage to use high-LET radiation in those labeling.
cases when the targeted cancer cells not are in
the cycle.
Characteristics of emitted
RADIONUCLIDES radiation
Energy Ranges
Different radionuclides are of interest and some Max Mean
considered for therapy are listed in Table 2. The Type of
interesting radionuclides can be divided in at MeV (mm) (mm)
least two groups: those that give long range decay
effects so that neighbors to the targeted cells can 90
also be damaged, and those that give short range Y β 2.3 12 4
effects so that mainly the targeted cells suffer. Ch
131
Among the long-range agents are high-energy I β 0.6 2.4 0.8 H
beta emitters such as 90-Y (max. range 12 mm) 211
At α 5.9 0.05 0.05 H
and 131-I (max. range 2.4 mm), suitable when 125
I Auger <0.03 <0.005 <0.001 H
the uptake is high but heterogeneous. Another
interesting nuclide is 32-P (max. range 8
mm).however, so far, it is mainly available in
the form of phosphate and therefore has been
considered too dangerous to use since
dephosphorylation and phosphorylation
processes might give unwanted incorporation of
the radioactivity in normal tissue such as bone.
The other extreme are the Auger electron
emitters, such as 125I, which have to be inside
the cell nucleus to give a significant radiation
dose to DNA. The range of most auger electrons
is only about 1-2 µm. Radioactive alpha
emitters, such as 211At, give alpha particles with
a range of about 50-70 µm and therefore belong
to an intermediate group.

The therapeutical efficiency of high-energy

beta-emitting radionuclides varies with the
tumor size and is highest at diameters
comparable to the range of the beta particles.
This has clearly been shown for 131I and 90Y by
Wheldon. It was shown that these nuclides are
not efficient for small cell clusters or single
isolated cells because the emitted electrons have
such high energy and range that they deliver
large fractions of their energy outside the small
cell clusters or single targeted cells. This is
shown in the recurrence probability curves
shown in Figure 10.
Figure 10 Tumor recurrence probabilities versus tumor
cell number when 131I (left) and 90Y (right) radionuclide
therapy is applied. The influences of type 1 heterogeneity
are indicated with dashed lines.
For single cells or small cell clusters, it is better
to target the tumor cells with short-range
radiation such as alpha emitters or agents other
than radioactive nuclides, such as toxins or
stable nuclides for neutron capture therapy. In
the latter case, 10-B can be applied because the
induced high-LET fission fragments have a

56
range of 6-9 µm, which is ideal to kill a single
targeted cell. The auger electron emitters can
only be used when there are reasons to believe
that the radionuclides are taken up in the cell
nucleus.
TARGETING PRINCIPLES
Several targeting principles for tumor selective
delivery of radionuclides to tumor cells have
been described in the literature. The most
popular so far is to tag the nuclides to
monoclonal antibodies either with more or less
specific methods or by attaching the
radioactivity to certain parts of the antibodies
such as the carbohydrate moiety. Antigenic
structures have been identified as potential
targets such as the membrane-associated form
of the carcinoembryonic glycoprotein antigen,
CEA, in colon carcinomas.
A new group of interesting targets are the
sometimes overexpressed growth factor
receptors, such as the EGF-receptor which is
overexpressed in several malignant gliomas,
adenocarcinomas and various squamous
carcinomas, and the PDGF-alpha receptor
which is overexpressed in certain gliomas. In
these cases, monoclonal antibodies with
specificity for the receptors and the
corresponding ligands loaded with radioactivity
could be applied. The latter is presently being
tried with EGF-dextran conjugates loaded with
radioactivity. Overexpressed signal pathways
and other related uptake mechanisms, such as
the mIBG (catecholamine precursor analogue)
uptake mechanism, can be used in certain cases,
e.g., for neuroblastomas.
HETEROGENEITY
The therapeutical response of any type of
radiotherapy is more or less heterogeneous and
the targeting processes give extra heterogeneity.
The heterogeneity problems in targeted
radiotherapy are of at least four different types:
Type 1 There might, in targeted therapy, be
heterogeneity in the expression of targets, or in
the uptake of the targeting substances, or both.
Antigenic expression and the uptake of
57
monoclonal antibodies can be heterogeneous,
which might also apply to other targets and
targeting substances. The heterogeneity
develops as a result of the karyotypic and
phenotypic instability which characterizes most
malignant tumors and which is most
accentuated in large tumors.
Type 2 If there is homogeneous uptake of the
targeting substance, there is heterogeneity in the
specific energy deposition. This applies both to
radionuclide therapy and external radiotherapy
and is most pronounced for high-LET
treatments where the dose is delivered through
few particle tracks.
Type 3 If a sub-population of cells is selected on
the basis of similar specific energy deposition,
then there is heterogeneity because not all
particle tracks give similar damage. Some tracks
might give no, or only a few, breaks in DNA
whereas other tracks might pass longitudinally
through a DNA molecule causing several breaks
and severe fragmentation. Thus, all cells with a
certain energy deposition do not suffer from
identical damage.
Type 4 If we select a subgroup of cells that all
have about the same specific energy deposition
and similar types of DNA-damage, then there
might be differences in the effects on cell
proliferation due to differences in repair. Some
cells might be near mitosis and there will be
only a few hours before the chromatin is
condensed and chromosome or chromatide
damage is expressed whereas other cells might
be in an early cell cycle phase or in a quiescent
state and therefore have long time available for
repair.
All four types of heterogeneity must be
considered in targeted therapy whereas types 2-
4 only apply in external radiotherapy. Types 2-4
also account for the "fractional survival" always
seen in radiation cell survival curves in vitro.
RECURRENCE
Considering typical cell survival values for
radiotherapy recurrence, probability curves can
be constructed for radionuclide therapy as
described by Wheldon. Such curves are shown therapy, somewhat higher values (about 1.5
above in Fig. 10. These curves show that the higher) can be applied.
recurrence probability is low at certain tumor
sizes and increases for both smaller and larger
tumors. Heterogeneity does not appreciably
change the optimal tumor size for treatment, but
decreases the total dose and thereby increases
the risk of recurrence and the curves are then
shifted upwards along the y-axis in Fig. 10.

WHOLE BODY IRRADIATION

Photons, which most often accompany alpha

and beta decays, are not of immediate interest
for targeting but they give an undesired
background dose to large areas of the body. The
most critical tissue is the bone marrow, which,
if the whole marrow is irradiated, sets the LD50
dose for humans to about 4 Gy. The bone
marrow syndrome is, after doses higher than 4
Gy, fully expressed after one month in humans,
which corresponds to the time it takes for most
blood cells and lymphocytes to die. The persons
then die because new blood cells and
lymphocytes have, due to the irradiation, not
been formed from the stem cells in the bone
marrow. A life can only be saved if bone
marrow transplantation can successfully be
carried out.

When whole body doses above 10 Gy are

applied, the gastrointestinal system shortens the
lifetime to about one week corresponding to the
time it takes for the gastrointestinal epithelium
to be degraded. The probability of saving a
human life after a total body dose higher than
10 Gy is zero.

In targeted radiotherapy, it is important to keep

the whole body dose, and especially the bone
marrow dose, well below 2-3 Gy. If it is not
possible to avoid this, preparations must be
made for bone marrow transplantation. In the
latter case, it might be possible to increase the
whole body dose up to about 7-8 Gy.
The dose values given in this section apply to
high dose rate situations (about 1 Gy/min),
commonly used in external radiation therapy. At
low dose rate (< 1 Gy/hour), which is the
common situation for systemic radionuclide

58
 CHAPTER 7
DETECTORS
Our five senses are limited and to understand
what really occurs around us we must
sometimes use devices -- detectors. Our
unassisted senses have limited means of
detecting ionizing radiation. Heavy ions
passing through the vitreous body of the eye
may produce light, which is perceived and ions
in the air present blurred perceptions of
"something". Roentgen discovered that his rays
gave rise to “phosphorescent" light when they
met certain material. Becquerel discovered
natural radioactivity by demonstrating that
mineral photographic plates become blackened.
The Curie couple learned to make use of the
electroscope to measure charges that radium
produced in air. So we have gradually learnt to
construct and use instruments that augment our
senses.
Using heat as a detector
Ionizing radiation transmits energy to the
irradiated material. Like all other energy
deposition, it gives rise to an increase in
temperature that can be used to monitor the
irradiation. The absorbed dose of 1 Gy (1 J/kg)
will increase the temperature by about 2.5 •10-4
oC in water, which is a fairly good equivalent to
tissue. Thus, irradiation with 4 Gy, which is a
fatal whole body dose, should then increase the
body temperature by only 0.001 oC. In radiation
protection, where the doses are in the range µGy
– mGy, the warming effect is accordingly quite
negligible.
Hence, measuring instruments in the laboratory
must be founded on other principles. They must
be sufficiently sensitive to give warning at dose
rates > 10 µGy/h. But they must also be able to
measure low radioactivity concentrations in
laboratory work or to measure radioactivity
distributions in Nuclear Medicine.
Varying measurement problems make different
demands on the instruments' performance. A
single instrument is seldom idealistic from all
viewpoints. Therefore, many different types of
instruments and equipment are developed. To
be able to interpret the measurements correctly,
we need to know the principles of the
59
instrument, the type of radiation it can indicate,
how it is calibrated and - not least - how to
handle the instrument.
Common types of laboratory and radiation
protection instruments and detectors will be
described in this chapter as well as their
characteristics and a range of applications.
Detection Principles
Most detectors for ionizing radiation use its
ability to cause ionization and excitation (see
Chapter 1). This chapter will just deal with
instruments based on this principle. Other
detectors may be based on the ability of certain
types of radiation to cause nuclear reactions
(e.g. in the detection of neutrons).
"Counts" of electrons
When matter is ionized a large number of
electrons are set free. Binding energies of outer
shell electrons are in the order of 3-6 eV, but
since the energy transfer is a statistical process
an excess of energy is needed. On average, in
most materials about 30 eV are required to
create one ionization. Energy not used for
ionization causes excitations, vibrations and
other atomic processes. A 1-MeV particle,
which deposits all its energy in a detector,
creates 30,000 primary, free, electrons. If all
electrons are collected and counted in a proper
way, we should be able to register not just that
the detector was hit by the particle but also the
energy, position and direction of the particle.
How much information we can get depends not
only on the detector principle but also on the
sophistication level of the detector system.
Materials that are insulators, such as gases and
certain solid materials, allow a direct count of
the ionized electrons. If a high tension is applied
no current will flow in an isolator. If the
material is irradiated, free electrons are
produced that can move affected by the electric
field (Figure 1a) and a measurable electric
current is produced (Figure 2).
If many particles hit the detector they may
contribute to a constant current at the level of a
few pA, which can be measured by an electrical
instrument. This current is just related to the
total energy deposited in the detector. No
information of the number or type of particles is
obtained.
If we instead measure the current from each
individual particle as a short current pulse more
information is obtained. We can count the
number of particles per second hitting the
detector, and we can integrate the pulsed current
and obtain information about how much energy
each particle deposits in the detector etc.
However, to measure current pulses, consisting
of a few thousands of electrons only, is
technically more difficult and puts other
demands on the detector material.
a b c can be distinguished chemically and the number
Excited or
ionized atom
At deexcitation
a light photon
Free electron
is emitted
with kinetic
energy
A changed
chemical state
Figure 1 Different effects of ionizing radiation.
a. Free electrons are produced by ionization. In
an electric field, an electric current or pulse is
produced which can be measured.
b. On ionization or excitation, a hole appears in
one of the electron shells. An electron from an
outer orbit fills this hole. A photon is emitted
with energy equal to the difference in binding
energies. For certain material, these photons
are emitted as detectable light photons.
c. The ionization changes the chemical state of
the material permanently. The number of such
changes can chemically be measured.
"Counts” of light photons
On ionization and excitation, a hole appears in
one of the electron shells around the atom.
Sooner or later, an electron from an outer level
60
will fill this hole and release energy that can be
emitted as a light photon (Figure 1b). Such
photons can be "counted" by light-sensitive
detectors (photo-multipliers or photo-diods).
The number of photons is proportional to the
number of ionizations and excitations in the
material. This is also related to the total energy
released in the detector by the ionization
radiation. Materials of this type are called
scintillators and may be gases, fluids or solids.
"Counts" of changed chemical states
Change in the chemical state occurs when an
atom or molecule undergoes ionization and
excitation (Figure 1 c). For example, if the
irradiated substance initially contains divalent
ions (e.g. Fe+ +) and an electron is knocked out,
trivalent ions are formed (Fe+++). These ions
of trivalent ions formed can be determined. The
number of ions formed gives a measure of the
energy that the radiation has deposited. Liquid
and solid material can be used in these types of
detectors.
particle
path
-
-
-
+
Figure 2 Principles of a gas detector. An
ionizing particle creates a number of ion pairs
in a gas between two electrodes. The electrons
move towards the positive electrode and the
positive ions towards the negative electrode. A
current flows through the detector and can be
measured either as pulses or in a dc-mode.
Most radiation protection instruments are
constructed on the basis of these three
principles: counting the number of electrons,
photons or changed chemical states. The
construction and use of the most common types
will be described.

Gas detectors
The principles of a gas detector are illustrated in
Figure 2. The factors that determine the detector
function are:
1. The voltage between the electrodes.
2. The design of the electrodes.
3. The composition and pressure of the gas.
4. The wall construction of the detector.
Influence of high tension
If no voltage is applied, the ion pairs, which
have formed, recombine (Figure 3a). This
means that the positive ions and the electrons
attract one another to reform neutral molecules
or atoms. No current is measured and hence, no
ionizing radiation is detected.
If a small voltage is applied produced ion pairs
can either recombine or are separated by the
electric field. The number of separated ion pairs
increases with increasing voltage, but is still
less than the number produced by the ionizing
radiation (Figure 3b).
Increasing the voltage further a stage is reached
where all the produced ion pairs are separated
and picked up by the electrodes (Figure 3 c).
The electrons and the positive ions are
accelerated in the electric field but collide
occasionally with neutral gas particles and lose
their kinetic energy.
a b c d e area
0 - - - -
0 + + + + N1 Number of ion pair
Figure 3 The influence of the voltage between
the electrodes of a gas detector.
a. If the voltage is zero, the ion pairs formed
recombine. No current flows through the
detector.
b. If the voltage is small, the ion pairs formed
can recombine or travel to their respective
electrodes. The current through the detector
61
becomes less than the number of primarily
formed ion pairs.
c. If the voltage is increased a little, the
primarily formed ion pairs move towards their
respective electrodes. The current measured is
equal to the number of primarily formed ion
pairs.
d. If the voltage is further increased the
electrons formed acquire such a high energy
that secondary ions are produced. The current
through the detector becomes greater than the
primarily formed ion pairs.
e. It the voltage is the same as in d, but the
primary events occur nearer the positive
electrode, fewer secondarily formed ion pairs
are created. The measurability of the process
depends on the geometry of the detector.
If the voltage is increased further (Figure 3 d
and e) the electrons will, between collisions,
gain that much energy that they, in turn, can
give rise to secondary ionization. This will
create ion pairs that were not produced directly
by the ionizing radiation. Figures 3 d and e also
show how the collected charge varies depending
on where the primary ionization took place. By
designing the electrodes suitably, detectors can
be constructed so that the collective charge is
practically independent of where the primary
ionization occurred.
I II III IV V
1012
Recombination area
Ion chamber area
1010
Number of collected ion pairs
Proportion counter area Geiger-Muller Spark area
108
106
104 N2 Number of ion pair
from a α - particle
102
from a β - particle
100
0 300 600 900
High voltage (V)
Figure 4 The number of collected ion pairs in a
gas detector as a function of the electrode
voltage. A detailed explanation of the diagram
is given in the text.
The influence of the voltage is summarized in
Figure 4 that gives the number of collected ion
pairs as a function of the increased voltage. The
diagram shows, as a comparison, the number of
primary produced ion pairs from a beta particle
(N1) and an alpha particle (N2) and how they
relate to the number of collected electrons.

l Recombination region
The number of collected ion pairs is smaller
than N1 and N2.

ll Ion chamber region

The number of collected ion pairs is equal to the
number of primary ion pairs N1 and N2.
lll Proportional region
The number of collected ion pairs is
proportional to the number of primary ion pairs
formed. The proportional factor, also called the
gas multiplication factor, can be as large as
100,000.
lV Geiger-Müller region
The number of collected ion pairs does not
depend on the number of primary ion pairs
formed. Even if the deposited energy varies, the
detector gives the same pulse height.
V Discharge region
A spark occurs between the electrodes for each
ionizing particle, which gives rise to ion pairs in
the detector.
Ionization chamber
A detector that works within the saturation
region in which all the formed ion pairs are
assembled is called an ionization chamber
(Figure 2) and is generally a portable
instrument. The ion flux produced in the
instrument can be measured directly.
Figure 5 A portable ionization chamber for
radiation protection use. The current measured
equals the primary formed ion pairs.
An ionization chamber with a capacity of 1 liter
exposed to an irradiation of 1 µGy/h produces
an ion flux of about 10-14 A. In a portable
instrument, it is difficult to amplify and measure
smaller currents. Hence, 1 µGy/h is usually the
lower measurement limit of such instruments.
By refining the electronics or increasing the
ionization volume, instruments with higher
sensitivity can be made but they become bulky
and cumbersome.
Proportional Counters
By increasing the voltage, primary produced
electrons can be accelerated to such a high
velocity that in turn they can cause further
ionizations.
particle
path
- -
-
+

Figure 6 A proportional counter. The

measured current is proportional to the number
of ion pairs formed. The ratio between
measured and primary formed ion pairs is
called the gas multiplication constant.

62
As is illustrated in Figure 3 d and e, the number
of assembled electrons depends on where the
primary ion pairs are formed. By shaping the
positive electrode suitably, this dependency on
position can be eliminated. If a thin straight
filament or loop is used as the positive
electrode, the field intensity close to it is so
great that it causes secondary ionization (Figure
6).
In the large volume of the detector, the primary
formed electrons migrate towards the positive
electrode without causing secondary ionization.
Only the electrons within the nearest
millimeters of the thin filament are exposed to
such high field intensity that they can cause
secondary ionization. Within this region, each
primary formed electron causes repeated
ionization, which results in intensification of the
primary current pulse. This ionization avalanche
can intensify the primary yield by a factor of
100 to 100,000. The phenomenon is called gas
multiplication. It is mainly the composition and
the pressure of the gas that determine the
sensitivity and the working voltage of the
proportional counter.
The size of the current pulse obtained from the
counting tube is proportional to the number of
primary ionization created by the penetrating
particle. This can be used in a mixed radiation
field to separate between particles, such as
alpha and beta particles, giving away different
specific ionizations. Such pulse screening
demands precisely stabilized high tension and
pulse height discriminators.
By working the proportional counter at
atmospheric pressure, the chamber can be
designed so that a weak radioactive sample can
be introduced into it. Thus a favorable
measuring geometry is achieved and at the same
time the method makes it possible to measure
particles whose energy is so low that they
cannot penetrate the chamber walls. This type of
detector is called a "flow counter". After
entering the sample, a suitable gas composition
in the detector is established by a slow flow of
counter gas (Figure 9 c).
63
Geiger-Müller counter (GM-tube)
If the high tension is increased above the
proportional region, the proportionality between
pulse size and primary ionization is lost. In
region IV of Figure 4, the so-called Geiger-
Müller tube's plateau, the detector pulses
become equally large for α- and β-particles. The
gas amplification can amount to 108 but the
pulse size does not vary very much. If the
voltage is increased still further, there is an
electrical breakthrough creating a spark.. We
then enter into the spark chamber area where
light or sounds from the sparks are counted.
In the GM-tube, the electron avalanche close to
the filament advances along the whole filament
(Figure 7) with a speed, usually 10 cm/µs. In the
gas cylinder around the filament, all electrons
are picked up by the electrode creating
positively charged plasma. This decreases the
electrical field gradient in the immediate
environment of the filament that is essential for
the genesis of an electron avalanche. When the
positive ions have diffused towards the casing
of the tube, the high electrical field intensity
around the filament is restored and the tube is
again ready to trigger a new event.
particle
path
- -
-
+
The area around
the central
electrode will be
competely ionized
Figure 7 Geiger-Müller detector. The discharge
around the filament produces so much positive
charge that the region of high electrical field
strength disappears. When the positive ions
have diffused towards the surface, the high field
strength returns and the detector is again
sensitive.
When the positive ions reach the wall of the
tube, they can knock out electrons from it and
thereby create new pulses unintentionally. This
can be prevented either by electronic
"quenching-circuits" that reduce the tube
voltage briefly (between 10-3 and 10-4 seconds)
or by making a gas mixture so that the tube
becomes "self-extinguishing".
As a rule, the gas mixture consists of an inert
gas such as argon at a pressure of 10 cm
mercury and a quenching gas, commonly
ethanol (10 %) or halogen (0.1%). Tubes
containing an alcohol admixture have a limited
lifetime (1010 pulses) since the alcohol
molecules are consumed, whereas halogen tubes
have the practical advantage of unlimited
lifetime.
Figure 8 This instrument contains a small GM-
tube. A larger GM-tube can be connected if a
more sensitive measurement is required. This
tube may be of the end-window type, equipped
with a very thin foil that allows low energy β-
particles to penetrate.
Gas detectors can be made in many different
ways for different uses. An instrument for
photon radiation with energy higher than 50
keV requires a thick wall around the gas (Figure
9a). Photons with high energy have very little
likelihood to interact with the gas itself. The
thick wall increases the probability for
interactions where high-energy photo- or
Compton electrons are formed. These can then
enter and ionize the detector gas.
Sample
Gas
inlet
Stright, cylindrical tube End window tube Gas flow tube
64
Figure 9 Different constructions of gas
detectors
However, low-energy photons and electrons can
not readily penetrate thick detector walls. For
these types of radiation, detectors with thin
walls must be chosen so that the radiation can
enter the detector gases. For low energy
electrons (e.g. beta particles from 14C)
extremely thin windows (1-10 µm) are essential.
Often a combination is used in that a thick-
walled detector is provided with a thin foil at
one side. Such a construction is usually called
an end-window counter (Figure 9 b).
To detect 3H-beta (energy < 18 keV) a
windowless detector (gas flow counter, Figure 9
c) has to be used. To achieve stable conditions,
the detector gas is allowed to flow at uniform
speed through the detector out by an opening
where the investigated object is introduced.
Radiation protection instruments that work
according to this principle have been
constructed to detect 3H but are generally
expensive and difficult to use.
The ionization chamber is principally the best
instrument (Figure 5) to measure the radiation
energy deposited or the dose equivalent. The
flow of ions through the chamber is directly
proportional to the absorbed energy.
Disadvantages are that the instrument is
relatively insensitive and can be heavy and
awkward to use.
The proportional counter gives a pulse that is
proportional to the energy, but it is not
especially suitable for measuring the absorbed
dose. When the proportional counter is used as
a radiation protection instrument, it is usually to
differentiate sparse and massive ionizing
radiation (e.g. α and β).
The Geiger-Müller counter gives no energy
information. However, by a suitable assembly
of the detector walls, the instrument can be
calibrated to show the dose rate equivalent
(µSv/h) for photons within a wide energy range
(50 keV - 3 MeV). It must be stressed that this
is only an approximate calibration and valid
only for photon irradiation within this energy
range. The great advantages of GM-detectors
are that they can be of robust make, easy to
handle, relatively sensitive, and cheap.
Scintillation Detectors
Many solid and fluid materials can convert the
energy of absorbed radiation into light. This
phenomenon, luminescence, has for a long time
been used in nuclear physics to demonstrate
particles and to make direct observations on
fluorescent screens, e.g., patients undergoing
radioscopy. As early as 1910, Rutherford used a
scintillation detector (zinc-sulfide) when he
made scattering experiment with α-particles. He
then observed the light flashes from the screen,
the scintillations, with his naked eye.
Today, sensitive light detectors (photo-
multiplier tubes and photo-diodes) can convert
transmitted light to an electric signal, which is
then recorded. The principles of a scintillation
detector are given in Figure 10. Requirements
for scintillating material to be used for detector
purposes can be summarized as follows:
1. High efficiency in converting energy in the
form of ionization and excitation to light. If a β-
particle deposits 100 keV in a scintillator with a
10 % conversion factor, only 10 keV is emitted
as light energy (photon of blue light ~ 3 eV).
Accordingly, at each scintillation about
10,000/3 ~ 3,000 light photons are obtained.
2. A scintillator must be transparent for the
light it produces.
3. The light photons must be emitted during a
short period (< 10 -5 s).
4. The frequency of the emitted light must be in
accordance with the light detector's response
function. A photo-multiplier tube usually has a
sensitivity of about 20 % to detect a blue
photon. For the example above, this means that
if 3000 blue photons hit the PM-tube only
3000*0.2 = 600 photons are detected.
65
Ionizing
radiation
Deposition of energy as
ionizations and excitations.
Material: Crystals, organic
liquids and gases.
A part of the energy is
transformed into light
emitted in flashes or
scintillations.
The scintillations are registered
Photomulti- by a light sensitive detector
plier tub which transforms the light into
electric signals.
The electric pulses are analyzed
and recorded by a special type
Electronics
of electronics, pulse electronics.
Figure 10 Principles of a scintillation detector.
Both organic and inorganic material can work
as scintillators. Organic scintillators can be
regarded as organic scintillating molecules in
solution, either in a liquid form or in a solid
phase. They usually have a density and an
elementary composition corresponding to the
body tissue. They are also characterized by great
speed, that is to say the light flashes are of short
duration. An advantage is that they can be made
in large volumes where several photo-
multipliers collect the light.
Inorganic crystals have greater density (2,000 -
8,000 kg/m3 ) and often have greater light yield
with longer after-glow time than organic
scintillators. The most common crystal in the
inorganic group, sodium iodine activated by
thallium, NaI(Tl) is available commercially in
sizes up to several dm3 and in thin slices with
diameters of 1 m or more. Due to favorable
characteristics, absorption, light emission, speed
and costs, NaI(Tl)-detectors have a great
significance in γ−radiation detection. These
crystals are very sensitive to humidity and are
therefore encased in an internally white,
reflecting wrapper with a window that permits
light to reach the photo-multiplier tube.
NaI(Tl) - detectors
Scintillation Radiation
The principle of a scintillation detector is shown
in Figure 11. For gamma detection it is an Scintillator
advantage if the scintillator contain high atomic
number materials, such as iodine, since it has a Photocathod
high probability to interact with gamma. A high
probability to absorb the total gamma energy is
also of advantage since it determines the
incoming gamma energy.

The produced light photons in the detector hit etc

the photoelectric cathode of the photo-
Dynods
multiplier tube. Generally, this consists of a
bivalent alkaline compound, e.g. SbKCs, which
has many loosely bound orbital electrons. Light
photons can expel these electrons in a photo
absorption process. The freed electrons are
accelerated in vacuum by an electric field Anode
towards an electrode. The kinetic energy
transferred creates more electrons, which in turn
are accelerated towards a new electrode and so R C
on. The electrodes are called dynodes and give -V+
rise to secondary electrons (compare gas
multiplication).

Typically, each dynode hit gives rise to 2 - 5
electrons. Generally a photo-multiplier tube has
6-10 dynodes and may yield an electron
amplification of about 106 times. Consequently,
for each light photon that produces a
photoelectron, 106 electrons are obtained from
the photo-multiplier tube. A 100 keV deposition
in the crystal may produce about 3,000 light
photons. Typically will this produce 600 photo-
electrons (20 % conversion efficiency) and in
the end a current pulse containing 600 · 106
electrons will emerge from the photo-multiplier
Figure 11 The principle of a NaI(Tl) detector.
A crystal of sodium iodide, usually in the form
of a cylinder, is attached to a photo-multiplier
tube with good optical contact. Other surfaces
are coated with reflecting material to force light
photons to hit the photo-multiplier tube no
matter where they are formed in the crystal.
X-ray from Photo peak
the K-shell
Back scatter Compton
Number of pulses/keV

1000
peak edge
tube.

The entire detector must be encased in a light-

and airtight covering to protect the photo-
multiplier from outside light and the crystal 100

from humidity.

10
0 200 400 600 800
Pulse height, γ-energy (keV)

Figure 12 Gamma energy spectrum of 137Cs

66
Generally, the sodium iodide crystal is "doped"
with a small amount of thallium (0.2 %) to
increase the light yield. Because of its high
density and the high atomic number of iodine,
the sodium iodide (thallium) detector is best
suited for the detection of gamma- and X-
radiation. When the pulse coming from the
detector is proportional to the deposited energy,
and the radiation frequently deposits its entire
energy in the crystal, the detector can be used
for energy analysis. A typical gamma energy
spectrum is shown in Figure 12.
Figure 13 Scintillator detector for radiation
protection purposes. There is either a thin
NaI(Tl)-crystal adapted for photon energies
from disintegration of 125I or a thick NaI(Tl)-
crystal designed for detection of higher gamma
energies.
Thin crystals make suitable detectors for
photons of low energy. The total crystal volume
is small and hence the efficiency for other
radiation, e.g. cosmic radiation, is small. Hand
instruments intended for the detection of 125I are
available and can be made sufficiently sensitive
to detect small amounts in the thyroid gland (a
few hundred Bq). At high photon energies, one
needs to increase the crystal volume to increase
the efficiency. Thereby the background
contribution from cosmic radiation increases
and these detectors become less suitable for
detecting radiation of low energy.
67
Plastic Crystal Detector
If the sodium iodide crystal is exchanged for a
plastic crystal, a detector is obtained that can be
suitable for both beta- and alpha- detection. The
light proof wall of the crystal can be made
extremely thin to let low-energy particles
penetrate.
This type of detector is also sensitive to photon
radiation but, due to its low density, not to such
a high degree as the sodium iodide detector. The
plastic crystal detector is not suitable for energy
analysis of photon radiation. The low atomic
number of plastic material makes the photons,
especially incoming photons, deposit only part
of their energy in the detector (Compton
interaction). The probability of photo absorption
is of course low.
Liquid Scintillation Detectors
A scintillator in the form of a liquid makes it
possible to mix the sample to be measured with
the actual detector material (Figure 14).
Solved
radioactive Liquid
sample scintillator
Photomultiplicator
tube “viewing” the
scintillations
Sample and Electronics counting
scintillator pulses and analysing
are mixed pule height
Figure 14 Principles for Liquid Scintillation
Counting. The advantage of this method is the
intimate contact obtained between sample and
detector. No walls prevent a direct transfer of
β-energy to the scintillator.
This technique maximizes the energy transfer
from sample to detector and hence is well suited
for measuring low-energy radiation from, for
example 3H, for which suitable radiation
protection instruments are lacking.
Figure 15 Liquid Scintillation Detector. The
instrument is very useful for radiation
protection work e.g. in contamination tests.
One common way to use the liquid scintillation
detector for radiation protection is to make a
sweep test. A bench surface or part of the body
suspected of being contaminated is washed with
absorbent material moistened in a suitable
solvent. The absorbed material is then put in a
scintillation flask (Figure 15), then put in the
scintillator and the sample is measured in the
detector. By analyzing the pulse heights,
information can be obtained as to whether the
material emits radiation energy.
Semiconductor Detectors
Solid material may be used as an alternative to
letting the radiation interact with a gas and
collecting the ions formed in it. Suitable
semiconductor materials are silicon and
germanium. Sometimes such detectors must be
cooled to a low temperature.
Semiconductor detectors that often have very
good energy resolution have frequently been
used for α- and β- spectrometry.
68
Photographic Emulsions
Ionizing radiation such as visible light can
affect the silver halide granules of photographic
emulsions so that in the development process
they can be reduced to silver. Thus, blackening
of photographic films may occur as for X-ray
photographic examinations. Photographic films
can also be used for dosimetry.
Figure 16 Two types of personal dosimeters. A
track film type is on the left. In films e.g. nitrate
cellulose films, high ionizing particles cause
tracks, which can be developed and counted. To
the right is seen a dosimeter based on
photographic film. When the film is irradiated it
gets blackened. The density can be measured to
give an idea of the dose of ionizing radiation
received.
Electrons or γ-radiation causes a general
blackening of film emulsions whereas heavy,
charged particles, which have proportionately
short range, give rise to trajectory tracks that
can be studied under the microscope. Such track
detectors may also be used for non-charged
radiation, e.g. neutrons. In the plastic film
backing, the fast neutrons are scattered against
the protons. In this process, low-energy protons
with high LET produce tracks that can be
measured and used to quantify the neutron dose.
In radiation protection work, film dosimetry is
of great importance for routine individual dose
measurements. Two emulsions of different
sensitivities are often used to ensure detection
in a wide dose range.
The density of the blackness is obtained by
using densitometers, instruments that record the
amount of light that passes through the film Free electrons
emulsion. The densitometer scale is graded to
give a direct reading of the density and dose. Ionizing radiation hit the
thermolumiscent material.
Bound electrons are lifted up
By protecting different parts of the film with in the conducting band.
different types of filters, one may separate Bound
electrons
between charged particles and photons. Low-
and high-energy gamma radiation can also to a
certain degree be separated. This is sometimes A certain proportion of the Free electrons
essential for estimating the risks associated with free elctrons are immediately Electron trap Light emitted
irradiation. falling back to the bound state
while emitting the excess energy Forbidden in an allowed
as a light photon. Other are transition transition
Thermoluminescence Dosimeters trapped in intermediate levels
with long life-time. Bound
electrons
In certain materials such as LiF, CaSO4, CaF2,
and BeO, the ionizing radiation can cause
excited conditions that can persist for a long The trapped electrons Free electrons
time. By heating the material, de-excitation can are lifted up to the conducting Electron trap Light emitted
band by heating the material.
be achieved whereby light photons are emitted They may then fall back to the Forbidden in an allowed
and can be recorded. The amount of light bound state emitting a light transition transition
emitted is proportional to the amount of photon which is detected.
Bound
radiation to which the dosimeter is exposed. electrons

The radiation transfer energy to the bound Figure 17 Principle of operation of the
electrons in the material. Some are energetically thermoluminescence dosimeter.
raised up into the conducting band (Figure 17a).
When these electrons fall back towards their
bound electron state, they may be trapped in an
excited level between bound and conducting
energy levels (Figure 17 b). These traps are
caused by impurities in the material. The
electrons cannot energetically fall down into the
bound energy levels since such changes are
"prohibited". Depending on the material, the
electrons can remain for months in these
"traps". The energy added in the heating process
during read-out lift up the trapped electrons into
the conducting level (Figure 17) from which
they can fall directly to the ground levels,
emitting light photons that are recorded.

The thermoluminescent dosimeter can to a

certain extent replace film as a personal
dosimeter and is also suitable for special
appliances such as finger dose measurements. It
is light, available in different forms, and easy to
tape securely to the fingers.

69
 CHAPTER 8
Scintillators in the laboratory
rather thick samples emit measurable amounts
SUMMARY of radiation. However, the detector material has
to be heavy and dense to detect high-energy
The following table lists different detector photons with some efficiency. The detector
principles for ionization radiation should also surround the sample to increase the
detection efficiency.
Type of Type of instrument Detector From such considerations two main detection
signals material techniques have developed to measure
Electrical Ionization chamber Gas radioactive biochemical or biological samples
pulses Proportional counter Gas • the liquid scintillation counter and
Geiger-Müller tube Gas • the gamma well counter based on NaI(Tl)
Semi-conductor Solid state Since such instruments are commonly used we
Chemical Film Photographic will here consider the technique in some detail.
Changes emulsions The measuring results depend on different
Chemical dosimeters Solids/fluids experimental conditions such as the type of
radionuclide used, the volume and the chemical
Prompt Scintillation counter Crystals or conditions of the samples, etc. Although
photons liquids modern instruments tend to automatically
Čerenkov counter Crystals or correct for a number of variables, untrained and
liquids non-experienced personnel may introduce
Delayed Thermo- Crystals severe measurement errors.
photons Luminescence
LIQUID SCINTILLATION COUNTING
Phosphorous Crystals
Imaging The main idea in this technique is that both the
Raising Calorimeter Solid or fluid sample and the detector are liquids, which are
mixed to have a close contact. The detector
of temp.
material converts the beta energy into light
photons.
DEFINITION OF THE PROBLEM
Sample Liquid scintillator
Most radionuclides of biological importance,
such as 3H and 14C, are pure, low energy beta
emitters. The maximum beta energy of 3H (18 Low energy beta will not penetrate
keV) has a penetration of about 10 µm in water. detector walls.
Solve the sample!
The mean energy of 6 keV yields about 200
Add liquid scintillator!
ionization events in total (30 eV per ionization). Shake!
Measure!
If we want to measure 3H with some efficiency
we can then conclude that Figure 1 The radioactive sample is mixed with
• the sample has to have negligibly mass or the liquid scintillator detector in a plastic or
thickness to avoid self-absorption glass vial (usually 2-20 ml in volume).
• the detector should not have an absorbing
window The vial with the sample and the scintillator is
placed inside a measuring chamber (Figure 2)
If a gamma- or X-ray-emitting nuclide, such as where all the emitted light is reflected into 2 or
125
I, is used the self-absorption in the sample is 3 photo-multiplier tubes.
of minor concern. The HVL of 125I-photons in
water is about 2 cm, which means that even

70
The detector Radioactive
decay
The energy transfer process from the radioactive Energy transfer
decay to the emission of light is described
Excited solvent
below. Chemicals that convert absorbed energy
molecule
to light are called “fluors”. Usually 5-10 gram Energy transfer
fluor is solved in one liter of aromatic solvent. N
The aromatic structure of the solvent is essential Excited fluor
O molecule
since it provides a first excited electron level
about 3-4 eV above the ground state. The
number of excited states is proportional to the Emission of
energy delivered by the beta particle. This light
excited level in the solvent has a rather long
lifetime but is sooner or later transferred to the Figure 3 A schematic figure of the energy
first excited level in the fluor (of somewhat transfer from the beta energy to the emission of
lower energy). This excited level in the fluor blue light photons.
has a short half-life (in the order of some
nanoseconds). In the decay a blue light photon Typically, about one blue photon is emitted per
is emitted. 200 eV deposited by the beta particle. The
detection efficiency for the PM-tube is about 20
MEASURING CHAMBER %. On average, it takes about one keV to create
one photoelectron as seen in Table 1. Thus, a
Reflectors Liquid scintillation vial
beta particle of 1 keV or less will probably not
give rise to a signal at all.
Photomultiplier Photomultiplier
tube 1 tube 2 Table 1 The table shows typical radionuclide
photon and photo electron yields for a single
decay event

Scintillation pulse Radio- Max Mean Typical Photo

nuclide Decay decay photon electrons
Background pulses Energy energy yield in the
(keV) (keV) PM tubes
Coincidence circuit. Accept simultaneous 3
H 18 5.6 30 6
pulses only. The summed output pulse is 14
proportional to the total energy delivered C 159 50 250 50
32
in the vial by the beta particle. P 1700 650 3300 660
Summed pulse
There is also a background to overcome. Now
and then, a spontaneous emission of an electron
Figure 2 A liquid scintillation sample is placed
from the cathode surface occurs (called the dark
inside the measuring chamber. The light
currant of the PM-tube). To distinguish the true
photons from the scintillation are forced to hit
signal from the background modern instrument
one of two photo-multiplier tubes. A coincident
use two or three PM-tubes coupled in
circuit sorts out background pulses coming
coincidence (see Figure 2).
stochastically from each PM-tube from true
events, which create simultaneous pulses in the
The solvent
two PM-tubes.
Pure benzene is a highly flammable liquid with
a high vapor pressure, which should be avoided
due to environmental reasons. Therefore, early

71
scintillators were usually based on toluene or
xylene. Modern scintillators are., from this
aspect, based on even better solvents (Table 2).
Table 2 Some data of commonly used solvents
in liquid scintillation detection.
Solvent Flash Equilibrium vapor
point concentration, 25 oC
(oC) (parts per million)
Toluene 4 36 000
Xylene 9 11 000
Phenyl-ortho-
149 1 400
xylyl ethane,
PXE
Di-isopropyl-
naphthalene, 148 1 600
DIN Scint
´HiSafe´
The high flash point solvents have several
advantages. They may be stored during normal
room conditions. The low vapor pressure lowers
the ventilation demands. These solvents have a
much lower penetration through the standard
polyethylene vials, which enables long term
counting. The health hazard is considered to be
lower than for toluene but contamination by the
skin should be avoided. A slight disadvantage is
that the counting efficiency is somewhat lower
than that for a toluene-based scintillator.
Water-based samples cannot directly be mixed
with organic solvents. Usually, a detergent such
as Triton X100 is introduced into the scintillator
liquid to cope with this problem. Small amounts
of water (up to 1.5 ml sample per 10 ml
scintillator) can then be solved in the
scintillator. If more water is added (5-10 ml
per10 ml scintillator) a stable gel is formed that
also can be used for counting. The intermediate
volume area, (1.5 -5 ml sample per 10 ml
scintillator) creates an unstable two-phase
system, which is not suitable for counting. The
borders between the different phases vary with
temperature, and chemical composition of the
sample, and details have to be taken from each
individual producer of the scintillation cocktail.
72
The scintillator
The solvents themselves are rather poor
scintillators since the decay time of the excited
states is rather long and the PM-tubes are not
sensitive to the wavelength emitted. A more
efficient scintillator, a fluor, with short half-life
and with a suitable frequency of emitted light is
therefore added. The most widely used primary
scintillator is 2,5-Diphenyloxazole, better
known as PPO (see Figure 4). It emits light with
fluorescence maximum at 365 nm.
N
O
PPO, 2,5-Diphenyloxazole
CH3 CH3
CH=CH CH=CH
Bis-MSB, 1,4-Di-(2-methylstyryl)-Benzene
Figure 4 Commonly used primary scintillator,
PPO and secondary scintillator, Bis-MSB.
A problem may arise if the sample absorbs light
in the frequency region of PPO emission. This
may be solved if a wavelength shifter
(secondary scintillator) is introduced, which
converts the 365 nm to a somewhat longer
wavelength. A common secondary scintillator is
1,4-Di-(2-methylstyryl)-Benzene (Bis-MSB).
Detection efficiency
As seen in Table 1 the number of photo-
electrons, i.e. the number of electrons emitted
from the cathode surface of the PM-tube, is few.
The coincidence condition requires that at least
one photoelectron is created in each PM-tube,
which corresponds to a deposited energy of
about 2 keV. In a continuous beta spectrum,
there will always be beta particles present with
such low energy that they do not trigger the
coincidence circuit. The counting efficiency is
thus always below 100 %. It is easy to realize
that this creates most problems in low-energy
beta spectra, such as 3H where the relative
amount of such particles is high. Typical
detection efficiencies for 3H are about 65 %, for
14
C 95 % and for 32P better than 99 %.
 CPM per energy 3H
14C
32P
18 159 1700
Energy (keV)
Figure 5 Energy spectrum of three typical beta-
emitting nuclides. The energy scale is
logarithmic to enable a simultaneously display
of the varying beta energy distributions.
The number of counts is often referred to as
counts per minutes (CPM) and is related to the
number of disintegration per minute (DPM) by
the detection efficiency ε as
CPM = ε * DPM
Another important parameter is the pulse height
since it is related to the total deposited energy.
14
C counting gives, in the mean, 8 times higher
pulses than 3H counting. This makes it possible
to separate between these two radionuclides in
dual labeling experiments. The common way to
display this is in an energy spectrum as seen in
Figure 5. Since there is a large difference in the
beta energies between different nuclides (a
factor of 100 between 3H and 32P) the energy
scale is usually logarithmic. The area of the
spectrum gives the total numbers of counts.
A common way to handle this is to amplify the
summed pulse (see Figure 2) with a logarithmic
amplifier. This pulse is analyzed by an analogue
to digital converter (ADC) and is presented by a
multi-channel analyzer (usually 1024 channels
or more). It is then easy to sum different parts of
the spectrum.
Quenching
In an ideal world the energy transfer in the
scintillation system would be both perfectly
efficient and undisturbed by environmental
factors. In the real word, however, energy losses
are substantial and depend on several factors.
Quenching is a term used to describe energy
73
losses in the energy transfer process (Figure 3).
There are three main types of quenching
processes that occur in the system:
• Physical quenching
• Chemical quenching
• Color quenching
Physical quenching
In gel counting, the scintillation system forms
small micro-droplets. The hydrophilic part of
the detergent is directed against the water
droplets whereas the lipophilic part is in contact
with the organic solvent. The diameter of the
droplets varies with the system parameters and
the amount of water in the scintillator, but is in
the range of 20-200 nm. If the radioactivity is in
the aqueous phase, the beta particles have to
penetrate a layer of water before they can start
to interact with the detector part. During this
water passage energy is lost that is not available
for light production.
Another example of physical quenching is
counting radioactivity absorbed in a solid phase,
e.g., a filter disc.
Chemical quenching
Any compound in the system that does not have
an aromatic structure similar to the solvent
interacts with the excited states. Such molecules
may steal the excited state and de-excite it in
small steps without creating a light photon. The
most severe quenching agents are halogenated
compounds (CCl4 > CHCl3 > CH2Cl2), ketones
and aldehydes. Less severe are salt solutions,
bases, acids, alcohol and water.
Color quenching
Color quenching occurs after the fluorescence
stage, when light-absorbing compounds
interpose. The number of photons that leave the
scintillation vial and hit the PM-tubes
decreases. The fluorescence emission takes
place in the blue region of the spectrum. A
sample collared in yellow preferably absorbs in
blue and therefore the order of severity of color
quenching is:

Red > orange > yellow > green > blue
In all types of quenching, energy is lost. The
pulse out is lower and the energy spectra in
Figure 5 are moved to the left. This means that
differently quenched samples are measured with
different counting efficiency. However, if the
quenching in an experiment can be proved to be
constant, then the CPM value may be used as a
substitute for DPM values.
Quench Correction
A methodology is therefore required to evaluate
the level of quench, determine the detection
efficiency, and correct the CPM result to give
DPM.
Quench correction adding an internal
standard
One method available is to add a known amount
of a radioactive isotope to the scintillator
sample mixture. This is called ''spiking''. Ideally
the isotope added should be in the same phase
as the sample. Using the following equation, the
efficiency of the energy transfer process can be
determined.
Efficiency = CPM increase/DPM added
The advantages of this method are that only
simple arithmetic is required to obtain a DPM
result. However, the disadvantages are many
and the method is only used in special
situations. The disadvantages are:
• Costly in terms of the amount of radioactive
isotope needed
• Costly in extra handling time.
• Costly in extra counting time (it is necessary
to count the sample before and after
spiking).
• Additional potential health hazard in extra
sample handling.
• Physical quenching can lead to errors in the
efficiency calculation.
• Increased errors due to multiple pipetting.
74
Sample Channels Ratio
To calculate the DPM of an unknown sample,
the counting efficiency of the sample must be
known. Spectral shifts are directly related to
energy losses and thus to counting efficiency. A
correction method, which uses the radioactivity
in the sample itself can be designed as follows:
 A number of calibration samples with
known amount of radioactivity are quenched
differently. The samples are measured
(CPM) and the counting efficiency (ε =
CPM/DPM) of each sample is determined.
 The spectral shift due to the quenching is
also quantified. The classical method is to
split the spectrum into two parts or two
''counting windows''. A ratio is formed
Count window 1
Sample Channel Ratio ( SCR ) =
Count window 2
 SCR is measured in all the calibration
samples and will vary with the degree of
quenching. We can now make a plot with
the efficiency (ε) on the y-axis and the
quench-measure (SCR) on the x-axis.
 For an unknown sample we can determine
SCR. From the plot we can then determine
the measurement efficiency of the unknown
sample.
Multi-channel analyzer (MCA) technology
where one can use thousand windows instead of
two to quantifying the spectrum shift of the
sample has improved this technique somewhat.
However, the main drawback is that one need to
have fairly high levels of radioactivity in the
samples to obtain good statistics in the quench
parameter.
External Standard Channels Ratio
Samples with low radioactivity content are thus
still a problem. The common way to overcome
this is to irradiate the samples for a short
moment with a strong, gamma-emitting source.
This irradiation produces Compton electrons in
the sample that will interact with the
scintillation solution as the beta particles do.
We can then measure a Compton electron
spectrum, which is shifted in a quenched sample
like a beta spectrum.
We can then create a quench measure, External
Standard Channels Ratio (ESCR), which can be
used to monitor sample efficiency in the same
way as above. It is independent of the sample
radioactivity and high statistical confidence is
achieved rapidly.
Modern equipment, which analyses the data
with MCA, can create special Quench
Parameters based on the external standard and
can from these measurements automatically
correct for various types of quenching.
CRYSTAL SCINTILLATION COUNTING
In liquid scintillation we measure the energy
deposition of charge particles. Due to the low Z-
number, gamma is detected with low efficiency.
However, all gamma-emitting nuclides also
emit charged particles such as beta particles,
conversion electrons, Auger electrons etc. and
can thus be effectively detected in a scintillation
detector.
So why use a gamma detector if you already
have a liquid? The answer is that gamma
counting is more simple and reliable. The
gamma rays penetrate even large samples so
there is no need to solve the sample. The sample
composition does not influence the
measurements and no quenching corrections are
needed. However, there is still a need to
understand the system in order to avoid pitfalls.
A gamma detector needs to be of high density
and to have a high Z-number to be efficient.
There are a number of crystal materials that
fulfill the following criteria:
1. High atomic number and density
2. High light output
3. High transparency
4. Fast decay time
5. Stable in time
6. Inexpensive
75
However, the most commonly used gamma
detector in a biological laboratory is by all
means a well counter based on NaI(Tl) (see also
Chapter 7). We will below discuss different
aspects of NaI-systems in some detail.
Sodium iodide spectroscopy
Because NaI(Tl) detectors have energy
resolution, they can discriminate between
gamma rays of different energies and hence can
map the energy spectra of radionuclides. Before
going into a detailed discussion of the features
of NaI(Tl) spectroscopy, it is useful to briefly
review the physics of gamma ray interactions
(see also Chapter 3). Over the range of energies
typically encountered in nuclear medicine and
biology procedures (30 keV to 1 MeV) there are
two gamma-ray interaction modalities:
photoelectric absorption and Compton
scattering.
In photoelectric absorption, the gamma ray’s
energy is completely absorbed by an inner-shell
atomic electron, The electron is ejected from the
atom with an energy equal to the gamma-ray
energy minus its atomic binding energy. The
empty vacancy left by this event is quickly filled
by outer-shell electrons with the emission of
characteristic X-rays and Auger electrons. The
cross-section (that is the probability that an
interaction occurs) for photoelectric absorption
depends on the cube of the atomic number (Z3)
of the absorbing materials. Because of the high
effective Z of NaI(Tl), photoelectric absorption
is an important interaction up to 1 MeV and is
the dominant interaction up to about 300 keV.
In low-Z materials such as soft tissue,
photoelectric absorption accounts for a small
fraction of photon interactions.
In Compton scattering, the incoming gamma ray
dissipates only a portion of its energy as it
scatters off a loosely bound electron. The energy
lost by the gamma ray is imparted to the
electron. The energy transferred to the scattered
electron depends on the scattering angle and is
at a maximum when the gamma ray is scattered
back in the direction it originally came from
(back-scatter). The cross section for Compton
scattering is only slowly varying with Z and
energy. As a result, Compton scattering is the
dominant interaction for low-Z materials (such
as soft tissue) and becomes increasingly
important in all materials as energy increases.
When the gamma ray energy exceeds 1.02 MeV
another interaction known as pair production
becomes possible. The gamma ray disappears
and is replaced by an electron and a positron.
The electron dissipates its energy and remains
stable. The positron dissipates its energy and
then captures an electron, resulting in mutual
annihilation and the emission of two 51l keV
photons (annihilation radiation). The cross-
section for pair production depends on the
square of the atomic number (Z2) of the
absorbing material and increases as the gamma
ray energy increases above the 1.02 MeV
threshold.
Photopeak
The most evident feature of the gamma-ray
spectrum is the photopeak, which results from
gamma rays whose energy is totally absorbed in
the crystal (Figure 6).
Total absorption
NaI-detector
Counts per channel
Photopeak or
Full value peak
Photon energy
Figure 6 Gamma spectrum of a NaI(Tl)-
detector. The shaded part corresponds to the
photopeak or full energy peak.
Because this is generally the result of a
photoelectric absorption, it is referred to as the
photopeak but any combination of interactions
that result in the complete absorption of the
gamma ray contributes to the peak. The fraction
of counts in the energy spectrum that fall within
the photopeak is referred to as the photo-
76
fraction and depends on the detector thickness
and the gamma-ray energy.
Compton features
It is possible for a gamma ray to be Compton
scattered in the detector and for the scattered
photon to escape without further interaction.
The energy deposited in the detector varies with
the scattering angle. This results in the so-called
Compton plateau, which represents the range of
possible energy deposition from Compton
scattering events (Figure 7). On the high-energy
side, there is a sharp drop off. This is the
Compton edge, and it corresponds to a gamma
ray that is scattered at 180 degrees in the
detector. The energy of the Compton edge is
calculated from Eγ -Eγ /(1 + Eγ/255) where Eγ
is the energy of the gamma ray (in keV).
Only a part of The gamma is
the energy stay scattered backward
NaI-detector NaI-detector
Counts per channel
Compton
edge
Distribution of
Compton electrons
Photon energy
Figure 7 Gamma spectrum of a NaI(Tl)-
detector. The shaded part shows the Compton
scattered energy deposit in the detector.
Escape peaks
When a gamma ray interacts with the sodium
iodine crystal, it is possible that only a portion
of the energy is absorbed. This is how the
Compton edge and plateau are formed (as just
described). Energy from interactions can escape
the crystal in other ways that yield discrete
peaks in the spectrum, so called escape peaks.
 Lead X-rays
Iodine x-ray leaves crystal
Lead x-ray is detected
NaI-detector
Counts per channel

Lead shield

NaI-detector

Counts per channel

Iodine Escape Peak Lead shield
Energy = Eγγ - 27 keV
Lead x-ray
Energy = 80 keV

Photon energy

Figure 8 Gamma spectrum of a NaI(Tl)- Photon energy

detector. The shaded part corresponds to a loss
of energy from the full energy peak due to an X-
ray escape.
For example, when photoelectric absorption
occurs, the characteristic X-rays from iodine are
emitted isotropically. Normally, these X-rays
are absorbed and contribute to the total energy
peak. However, if the interaction takes place
near the edge of the detector, the X-rays may
leave the crystal. This creates an additional peak
called the iodine escape peak with a pulse
height 27 keV less than the total gamma-ray
energy (Figure 8). Iodine escape peaks are
usually only evident for low-energy photons (<
100 keV), in which a substantial fraction of the
gamma rays are absorbed near the crystal
surface.
Another type of escape peak may be observed
when high-energy gamma rays are counted and
Figure 9 Gamma spectrum of a NaI(Tl)-
detector. The shaded part corresponds to an
incoming lead X-ray, which is created by
Compton scatter in the detector shield.
Because most detectors are shielded with lead,
it is common to see a peak corresponding to the
lead X-ray energy (approximately 80 keV). This
is emitted when the gamma ray undergoes a
photoelectric absorption in the lead shielding
and the lead characteristic X-ray is absorbed by
the detector (Figure 9).
Coincidence sum peaks
Two gamma in the
same decay are
detected simultaneously NaI-detector
Counts per channel

pair production is possible. If the annihilation

takes place in the crystal, one of the 511 keV
annihilation photons, or both, may escape the
crystal. Thus two additional peaks can be seen, Coincident
Sum Peak
one at 511 keV below the true photopeak and E=Eγγ1+Eγγ2
the other at 1.02 MeV below. It should be noted
that if pair production occurs in the vicinity of Eγγ1 Eγγ2
the detector (for example, in the lead shield), a
peak at 511 keV also shows up in the spectrum.
Photon energy
Figure 10 Gamma spectrum of a NaI(Tl)-
detector. The shaded part corresponds to the
summation of two simultaneous gamma-ray
energies.

77
If two gamma rays or X-rays are absorbed in the
crystal within a short time, the magnitude of the
scintillation corresponds to the sum of the
photon energies. This produces a peak on the
spectrum whose apparent energy is equal to the
sum of the two absorbed photons and is referred
to as the sum peak (Figure 10).
Coincidence sum peaks can occur from a
variety of situations: multiple gamma-ray
emission in a cascade (111In), emission of a
gamma and characteristic X-ray in electron
capture (125I) or counting a high-activity source.
Coincidence sum peaks are much more likely in
well-counter geometry where the gamma rays
and X-rays have a much higher probability of
being simultaneously detected.
Characteristic X-rays
Radionuclides that decay by electron capture
necessarily emit characteristic X-rays and Auger
electrons because of the inner-shell vacancy that
results from the capture. As a result, there is a
peak on the spectrum corresponding to the
absorption of these X-rays (Figure 11). This
peak is often very prominent especially when
the rate of gamma emission is low. An example
of this is 201Tl, in which the 68- to 80-keV X-
ray peak is the most obvious feature in the
spectrum.
Gamma and characteristic x-rays
are emitted in the decay
NaI-detector
Counts per channel
Characteristisc
x-ray Peak
Photon energy
Figure 11 Gamma spectrum of a NaI(Tl)-
detector. The shaded part corresponds to the
absorption of a characteristic X-ray emitted in
the decay.
78
Backscatter peak
When a radioactive source is located some
distance from a detector and has material behind
it, a distinct peak is evident on the spectrum that
is due to gamma rays that have been Compton
scattered at 180 degrees and then totally
absorbed in the detector. These gamma rays
initially are directed away from the source but
are, after being backscattered, detected (Figure
12).
The reason for a distinct peak is that gamma
rays scattered at other angles cannot reach the
detector. Backscatter peaks are most intense
when the material behind the source is dense
and has a high Compton cross-section. The
energy for backscatter peak is Eγ /(1 + Eγ /255),
where Eγ is the energy of the gamma ray (in
keV).
Media scatter
If the gamma rays travel through any material
on their way to the detector, Compton scattering
may occur and the scattered gamma rays may be
detected (Figure 12). These scattered gamma
rays span the range from the backscatter peak to
primary gamma-ray energy for single
interactions. If the medium is thick enough,
multiple interactions are likely to occur, and the
scattered photons will cover an even wider
range. Compton photons produced outside the
detector and in the detector are not possible to
separate. As a result, when it is important to
discriminate scatter radiation, only photopeak
events are considered.
 energy peaks are broadened as shown in Figure
13.
NaI-detector
OBJECT Peak maximum counts
or NaI-detector
SHIELD

Counts per channel

Counts per channel

Forward
Backward Half maximum counts
spread Compton γ
Compton γ

∆E
E
Compton electrons
from the detector Photon energy
Photon energy
Figure 13 Gamma spectrum of a NaI(Tl)-
Figure 12 Gamma spectrum of a NaI(Tl)- detector. The figure illustrates the way to
detector. Scatter from the detector shield or quantify the energy resolution of a detector.
from an object is scattered into the detector.
Most scatter have energies from zero to the Energy resolution is quantified by determining
energy of the gamma. However, backscatter the amount of spread with respect to the
(photons scattered backwards)l have a more gamma-ray energy. The spread of the peak is
defined energy which appears as an extra peak measured in kiloelectron volts at the half-
in the spectrum. maximum level (full-width-at-half-maximum,
FWHM). Thus, energy resolution is defined as
GENERAL PROPERTIES OF NaI(Tl) Energy resolution = 100% x ∆E/E =
SYSTEMS
=100% *FWHM (keV)/Eγ(keV)
Although NaI(Tl) counters can be configured
Energy resolution improves (gets smaller) as the
and used for a number of different purposes,
photon energy increases. The 662-keV gamma
they have some common characteristics
ray from 137Cs is often used to measure the
including energy resolution energy calibration
energy resolution of NaI(Tl) detectors, and 7%
energy linearity, detection efficiency and count
is a typical result.
rate performance. These will be detailed in the
following sections.
Calibrating a spectrometer
Energy resolution
Because of the proportionality that exists
between the pulse height of the detected signal
Gamma rays are monoenergetic. A detector with
and the energy absorbed from the gamma-ray
perfect energy resolution would generate pulses
interaction, it is possible to calibrate the
with the same pulse height for each gamma ray
equipment in terms of energy. Typically, the
that was totally absorbed and the energy
pulse height analyzer (PHA) has a multi-turn
spectrum for these pulses would be a single line.
dial for adjusting the baseline (lower level) and
Sodium iodide detectors do not have perfect
another dial for adjusting the energy window. In
energy resolution because of statistical
many cases, it is useful to calibrate the PHA
fluctuations in the number of electrons liberated
(MCA) to a true gain of; that is, where 1 unit on
at the photocathode of the PMT. As a result, the

79
the baseline dial corresponds to exactly 1 keV
of energy. This can be done as follows:
1. Take two radionuclides with different
energy gamma rays such as 131I and 99mTc.
The gamma-ray energies should be neither
too close in magnitude nor too far away.
2. Adjust the baseline and window settings for
a 10% energy window centered on the
higher energy gamma ray assuming a true
gain of 1. For the 364-keV peak of 131I, this
would correspond to a window setting of 36
and a baseline setting of 346.
3. Put the high-energy source in front of the
detector. Adjust the gain of the amplifier or
the high voltage to maximize the detector
count rate.
4. Readjust the PHA (MCA) for a 10%
window on the lower energy gamma-ray
energy, again assuming a true gain of 1. For
the l40-keV peak of 99mTc this would
correspond to a window setting of 14 and a
baseline setting of 133.
5. Put the low-energy source in front of the
detector and verify that the count rate is
maximized. If it is not the amplifier gain
and the high voltage must be readjusted.
Steps 3, 4 and 5 should be repeated until
maximum count rates are achieved for both
radionuclides at their respective settings.
With the detector set at a true gain of 1, the
appropriate settings can easily be calculated
if it is necessary to change the coarse gain
on the amplifier. For example, if there is a
true gain of l and the coarse gain is
increased by a factor of 2, the baseline
settings are all multiplied by 2. In this
instance, a 20% window centered on 99mTc
has a window width of 56 and a baseline of
252.
Energy linearity
The pulse height of the scintillation signal is
proportional to the energy deposited in the
gamma-ray interaction. This statement is not
true over a wide range of energies because of
non-linearity in the spectrometer. For example,
if the spectrometer is set to a true gain of 1, it is
unlikely that both the low-energy photons (such
as the 27-keV X-rays from 125I) and high-energy
80
gamma rays (such as the 662-keV gamma rays
from l37Cs) will fall exactly at their expected
settings. Although this is not a problem when
setting up the spectrometer for gamma rays, that
are relatively close in energy, it must be
considered when using radionuclides whose
emissions fall at the extremes of the energy
range.
Detector efficiency
Not all gamma rays emitted from a source are
detected. Some of the gamma rays never reach
the detector and of those that do, not all interact.
The detector efficiency quantifies the fraction of
gamma rays that are detected. This can be
considered from two points of view: the fraction
of photons hitting the detector or the fraction of
photons emitted from the source. The first
depends on the intrinsic efficiency of the
detector whereas the latter depends on both the
intrinsic efficiency and the source detector
geometry.
Geometric efficiency
The intensity of gamma rays emitted from a
point source obeys the inverse square law:
I(r) = I(0)/r2
The inverse square-law is a natural consequence
of the fact that gamma rays travel in straight
lines. Imagine that there is a point radioactive
source at the center of a sphere. The density
(gamma rays per square centimeter) at the
surface of the sphere is I(0)/4πr2 where I(0) is
intensity of the gamma rays emitted from the
source and 4πr2 is the surface area of the sphere.
Suppose we have a detector located on the
surface of the sphere at a distance, r, from the
source. The fraction of gamma rays that hit the
detector is directly proportional to the area that
the detector covers on the surface of the sphere.
The geometric efficiency is defined as
gp = (Area of detector) x cos(φ) / 4πr2 Eq(1)
where gp is the geometric efficiency for a point
source and φ is the angle between the surface
and the detector. If the detector is pointed
directly at the source φ = 0 and cos(φ) = 1. The
geometric efficiency can be improved by either
increasing the area of the detector or decreasing
the distance between the source and the
detector. For scintillation detectors, geometric
efficiency is maximized with a well design, in
which a channel is bored into the surface of the
NaI(Tl) crystal. The source is placed inside the
well and is surrounded in nearly all directions
by the detector. It should be noted that Eq(1) is
not appropriate for very small source-to-
detector distances. When the diameter of the
detector is large compared with the source-to-
detector distance, then the appropriate equation
is gp = ½(1 - cos(φ)) where φ is the angle
between the source and the edge of the detector.
For example, if the source is sitting right on the
detector, then φ = 90 degrees and gp = 0.5. If the
source is positioned in a well chamber in which
φ = 160 degrees, then
gp = ½(l - (-0.94)) = 0.97
Because the attenuation coefficient for NaI(Tl)
decreases rapidly with gamma-ray energy, the
thickness of the detector must increase to
maintain intrinsic efficiency. In Table 3, the
thickness of NaI(Tl) required for an intrinsic
efficiency of 0.9 is calculated at selected
energies. The table also includes the thickness
required for a photo-peak efficiency of 0.9.
WELL COUNTER
A well counter is a NaI(Tl) detector that has
been specially designed to maximize efficiency.
As shown in Figure 14, the channel (or well) is
made in the center of the crystal. With this
design, the source can be placed inside the well
to obtain a high measuring efficiency. The
intrinsic efficiency can be high since large
crystals (5 x 5 or 7.5 x 7.5, diameter x height in
cm) can be used.

Intrinsic efficiency

The intrinsic efficiency (ε) is the fraction of

photons that hit a detector and are detected. It is
determined by the thickness (t) and the linear
attenuation coefficient (µ) of the detector
material, ε = 1 - exp(- µt). The intrinsic
efficiency is high (that is, close to 1) when the
product µt is large, thus we want either µ, or t,
or both to be large.

Table 3 Sodium iodine intrinsic efficiency of

some selected gamma energies. A gives the
thickness needed to have 90% intrinsic
efficiency of any energy deposition in the
crystal. B gives the thickness needed to have
90% intrinsic efficiency of total gamma ray
energy absorption in the crystal.

Gamma Thickness of Thickness of

energy (keV) NaI(Tl) (mm)A NaI(Tl) (mm)B Figure 14 A schematic view of a well counter
30 0.5 0.5 for gamma counting. The well, i.e. the hole in
140 5.5 9 the crystal, may be different in commercial
172 8 14 instruments due to how the samples are
247 20 40 changed automatically. It is easier to arrange
364 31 50 this technology in a through-hole detector but
511 52 69 the counting efficiency is somewhat lower due
662 52 84 to the larger hole.

81
 approximately 10% losses when the total count
Well counters that are used routinely to assay rate is 20,000 cps. If a high-energy sample is
large number of samples are usually automated. being counted (for example 18F) the photopeak
In these devices, the samples to be assayed are count rate at which these losses appear may be
loaded into counting tubes that sit in special below 10,000 cps.
racks. A transport mechanism allows each
sample to be counted sequentially for a pre- Sample volume
selected time interval, and the results are stored
as a computer file. It is possible to accurately The volume and geometry of the samples being
assay the absolute activity of samples in a well assayed should be kept constant for two reasons.
counter. This, of course, requires a calibration
source to relate the detected counts to
radioactivity units. It also requires knowledge of
the problems presented by count rate losses,
background correction, and the sample
geometry. These problems are briefly discussed
in the following sections.

Figure 15 A commercial gamma counter

Dead time losses

The finite temporal resolution of the NaI(Tl)

detector results in count losses. These become
increasingly important as the total count rate
seen by the detector rises. Count rate losses are
a particular concern when assaying samples that
span a wide range of activities. For
radionuclides with short half-lives, the best
alternative is to wait until the samples decay Figure 16 Illustration of sample volume and
down to the point when the losses can be detector geometry for optimal and reproducible
ignored. It should be noted that in many well counting.
counters, the count rate at which dead time
losses are manifested could be relatively low, First, the detection efficiency depends on the
especially for high-energy radionuclides that source distribution in the well.
have a small photo fraction. For example, if the
pulse width at the amplifier is 5 µsec, there are

82
Second, the amount of self-absorption within
the sample also depends on the source
distribution.

It should also be noted that absorption of low- 0.4

energy gamma rays is highly dependent on the 0.3
N = 20
p = 0.05 p = 0.15 p = 0.5
material and wall thickness of the source
0.2
containers. Therefore, if assays from a large
number of samples are made, care should be 0.1
taken to ensure the uniformity of the source 0
containers as well as the source volumes 0 5 10 15 200 5 10 15 200 5 10 15 20
r, the number of outcomes in N tries
Background
Figure 17 The binominal distribution
Background refers to detected counts that do not illustrated for some values of the number of
originate from the desired sample. Because of tries and probability for one of the outcomes.
the high sensitivity of NaI(Tl) detectors for
gamma rays, they must be heavily shielded to The distribution P(r) is illustrated in Figure 17
reduce the counts from sources outside the well. for some values of N and p. In a statistical
Usually, several centimeters of lead surround distribution the sum over the number of
the detector. If many samples are to be counted outcomes should be one.
over a long interval, several background checks
should be made just in case there is a time-
dependent factor associated with the
background.

Dynamic range Important parameters in a statistical distribution

are the mean and the variance and they can for
The magnitude of activities that can be this distribution be expressed as
accurately measured with a well-counter range
from approximately 20 Bq to 20 MBq. On the
lower end, the limitation is the uncertainty in
the background count rate, and the upper end is
limited by dead time losses.

COUNTING STATISTICS If we consider the radioactive decay the

The radioactivity decay can from a statistical
point of view be regarded as tossing a coin.
There are two outcomes since during a time
interval, either the nucleus decay or it does not.
Events of this type have statistically a binominal
distribution. The probability to decay can be set
to p, and the probability not to is then 1-p.
In N tries, e.g. N radioactive atom are
investigated during the time interval, r atoms
may decay. The relation between the number of
tries and the probability gives a distribution of
the number of outcomes, which can be
described by the relation below
probability for decay, p, is small. During one
second, a nucleus changes its conditions 1010
times or more, but may still not be able to
decay. However, the number of tries, i.e. the
number of nuclides, is also large, 1010 or more.
We know that each nuclide has a specific decay
constant, which somehow is a measure of this
nuclide’s probability to decay. If we in the
binominal distribution let N --> ∞ and p --> 0
while we are keeping N*p= constant = µ we
obtain the following relations

83
This relation is called the Poisson distribution
and is said to be the statistical distribution for Table 4. Counting statistics
rare events, such as the radioactivity decay. The
distribution has the mean µ and the variance of Number of One standard Relative error
the distribution is also equal µ. In fact, the only Counts, N deviation, N N /N as %
variable in the distribution is the mean value µ. 100 10 10
If we vary this value, the distribution varies as 1000 32 3.2
shown in Figure 18. 10000 100 1
0.4 100000 320 0.32
λ=1
0.3 t=1 t=3 t=10
Since we are working with a Gaussian
0.2
distribution we can apply all knowledge about
0.1 error propagation in this type of distributions.
0 The only thing to remember is that the variance
0 5 10 15 200 5 10 15 200 5 10 15 20 is equal to the mean, which simplifies the
r, the number of outcomes calculations.

Figure 18 The Poisson distribution shown for If we have two measurements, N1 and N2,
some values of the distribution parameter µ. obtained during the same time, we obtain the
following relations for the error propagation in
For low values of µ, the distribution is rather addition and subtraction
skew but with increasing value of µ it becomes
more symmetric. In fact, it is possible to show N1 + N2 ± N1 + N 2
that for large values of µ the distribution
converts into a Gaussian distribution.
N1 - N2 ± N1 + N 2
However, this Gaussian distribution is special
since the variance is equal to the mean of the The relative errors are 1/ N1 and 1/ N 2 . If
distribution. This can be used in nuclear we divide or multiply the data, we use the
counting to estimate the counting statistics from relative error the following way
one single measurement.
Consider the following example. We measure M = N1 x N2 or M = N1/N2
one sample with a nuclear detector and obtain
the number of counts N. The sample or detector ∆M/M = 1 / N1 + 1 / N 2
is of little interest and so is the measuring time.
Only the number of obtained counts is
A common situation is that we would like to
important. We know that the measurement is
subtract the background from the sample and to
restrained by the Poisson statistics. The Poisson
calculate the error. If the count rate is 300 CPM
distribution can be approximated with a
for the background and 600 CPM for the sample
Gaussian distribution (if N > 25) and with the
and we measure the two values for one minute,
mean = variance. Large values of N twill give a
we obtain Ns = 600 and Nb = 300. We then
rather narrow distribution ´, which means that N
obtain
is a good approximate value of the mean and the
variance i.e. µ≈N and σ2≈N. The error in the
M = Ns - Nb = 300 ± 600 + 300 = 300 ± 30
measurement can then be written as N ± N.
If we increase the time we measure the
One must remember that it is an approximation. background to 100 minutes, we obtain
However, it enables us to estimate the error of
Nb=30000 ± 173 and
the measurement from one single value.

84
M = Ns - Nb/100 = 300 ± 600 + 1.73 * 1.73
or M = 300 ± 24.6

Another example illustrates the error

propagation if a variable does not follow
Poisson statistics but normal Gaussian statistics.
We measure a sample and get N = 6000 CPM.
We would like to know the absolute value of
the radioactivity and the uncertainty in the
determination. For the detector we need two
values, the efficiency which is 50% and the
relative error (standard deviation) of this
determination, which is 5%. We then obtain

A = N / 0.5 = 6000/60/0.5 = 200 Bq and

∆A/A = 1 / 6000 + 0.05 * 0.05 = 0.052

A = 200 ± 10.3 Bq

85
 CHAPTER 9
NUCLEAR BIOLOGY AND MEDICINE: NUCLIDE PRODUCTION
GENERAL ASPECTS
Nuclides are divided into two groups: stable and
unstable, i.e. radioactive. The number of known
nuclides is more than 2000, of which 280 are
stable and the remainder radioactive. Half-lives
vary from a millionth of a second to millions of
years. Most radionuclides are very short-lived.
Radionuclides undergo transformations or
decay, emitting α, β−, β+, γ, X-rays, neutrinos,
conversion electrons, Auger electrons, protons,
neutrons and heavy ions.
The use of nuclides, both stable and radioactive,
has steadily grown in importance in science,
technology and medicine during the last 70
years. If we restrict ourselves to the area of
biology and medicine, the dominant use is the
tracer technology, i.e., to label bio-molecules to
follow their chemistry, kinetics, metabolism and
catabolism in vitro and in vivo. The application
areas are in fundamental science such as
investigating new biochemical pathways as well
as diagnostics and therapy. Other areas of
application are external and intra-cavity therapy.
Isotopic labeling
2
Stable nuclides of biogenic elements such as H,
13
C and 15N and even more radioactive nuclides
such as 3H, 14C, 32P, and 35S have played an
important role in biology. The reason for this is
simple. Important biomolecules can be labeled
by exchanging an adherent common, stable
nuclide, i.e. 12C, to a stable non-common
isotope, such as 13C or a radioactive isotope of
the same element, i.e. 11C or 14C. This is
referred to as isotopic labeling. The small
change in weight of the two isotopes is usually
negligible and from a chemical and biomedical
point of view, they are identical. The labeled
molecule can be administered as a tracer in
amounts and in such a way that it does not
disturb the normal metabolic pathways.
However, some care must be taken. If one
replaces H with 2H or 3H, the change in weight
is dramatic. Usually, this does not matter but if
the labeling position is critical e.g. close to a
chemical bond cleaved by an enzyme, the
86
enzymatic function may be slowed down
considerably.
The use of stable nuclides has advantages
especially in vivo, since no radioactivity and
associated biological hazards are involved.
However, sensitivity is limited due to a natural
background. The detection of natural isotopes is
also somewhat more technically complicated
and expensive, whereas radioactive nuclides can
be detected more easily and at lower
concentrations. Examples are auto-radiographic
techniques that have provided regional
information down to the sub-cellular level and
at concentrations of pM.
Non isotopic labeling
However, in Nuclear Medical diagnostic
procedures there wass a problem. Very few
radionuclides of biogenic elements emit suitable
γ rays that could easily be detected outside the
body. Instead one had to rely on radioisotopes
of non-biogenic elements, such as 67Ga, 99mTc,
111
In and 131I. They had the right physical
properties, were easily available and could
easily be labeled to biomolecules.
However, small molecules, such as amino acids,
were found to lose their specific biological
activity when labeled with such nuclides. 125I-
thyrosine could be transported across the cell
membrane but could not be incorporated into
the proteins, i.e. the membrane transport system
was fooled but not the more specific tRNA-
system. Other molecules, such as 125I-IUdR,
could mimic the biogenic molecule thymidine
and were incorporated into the DNA-molecule
but at a lower rate. In this case the iodine
replaces a methyl-group, a quite dramatic
change. However, the iodine atom has about the
same van der Waal radius as the methyl-group,
which is enough to trick the biology.
Labeling with analogs
Elements of the same group in the periodic
system may to some extent replace each other in
the biological system. Strontium can exchange
calcium in the bone metabolism and rubidium
can replace potassium. Even more heavy
elements were found to work, such as 201Tl+, which use higher photon energies (511 keV) but
which also replaces potassium and is a in coincidence (Positron emission tomography,
commonly used radionuclide in heart studies. PET). The labeling of the biomolecule should
Thallium is a well-known poison but small keep the biological activity and the kinetics of
amounts, well below toxic levels, are given in the labeled molecule intact. The label should be
the clinical investigations. In fact, the amounts stable in vivo for the time of the investigation,
given are even below the levels contained in and the radiation dose to the patient should be
foodstuffs in the market. small. In therapy, about the same conditions
should be fulfilled but the radiation dose should
The approach to use analogs to label be high in the tumor but small in normal tissues.
biomolecules was tried. One found, for
example, that selenium could replace sulfur in The different aspects of radioactive labeled
biomolecules e.g. 75Se-methionine had about compounds are illustrated in Figure 1. Different
the same metabolism as biogenic methionine. nuclides have to be produced both in an
optimal, efficient and economical way.
Radionuclide therapy Different production routes give a product that
might be useful in some circumstances, but not
Isotopes of iodine, 125I and 131I, were suitable all. In order to understand these problems, it is
for non-isotopic labeling of a variety of necessary to have an understanding of all the
biomolecules. However, iodine is also a underlying techniques, their advantages and
biogenic element but only in a very special shortcomings.
organ, the thyroid. The natural high uptake of
iodine in thyroid (30-50 % of administered Physical aspects
iodine) was early used, not only for diagnostic Radionuclide Biochemical aspects
• type of decay
purpose but also for therapy in malignant • half-life Antigen
TUMOR
thyroid tissues as well as in metastasis. A • production • tumor specificity
• distribution after • tumor density
suitable amount of 131I (about 1 GBq) gave a MAb catabolism • binding availability
sufficiently high radiation dose to kill the Imaging device Antibody
• radionuclide
tissues taking up iodine but with minor effects • resolution
• specificity
• affinity
to other tissues. One may consider this as the • sensitivity • size
• availability • pharmacokinetics
first use of “targeting” therapy. The same • catabolism
situation was found in bone metastasis, where Clinical aspects Radiolabeling method
the calcium analog 89Sr could be used for Diagnostic value
• efficiency
• effect on antibody
• tumor to tissue contrast
therapeutic treatment of severely spread • cost/benefit
• in vivo stability
malignant bone diseases. Side effects ANTIBODY
• dosimetry
• HAMA
When the first tumor selective molecules were
developed during the 70s (Mabs) one had a
great hope to use the specific uptake in the Figure 1 Different important aspects when
tumors for tumor treatment with radionuclides. choosing the optimal condition for a nuclear
The technique has found some applications but medical application.
is still under development. Smaller molecules
such as tumor receptor specific peptides and RADIONUCLIDE PRODUCTION
hormones are today used for the same goal.
There are two major ways to produce radio-
Different applications set different demands on nuclides: using reactors or particle accelerators.
the radionuclide used. A radionuclide used for We can say that we irradiate either with
diagnosis should emit photons in the range 100 neutrons (thermal, slow or fast) or with charge
-200 keV in order to be optimal with today’s particles such as protons, deuterons, alpha or
common detector, the Anger gamma camera. heavy ions. Since the target is a stable nuclide,
Other detection modalities have been developed we generally obtain either a neutron-rich

87
radionuclide (reactor produced) or a neutron-
deficient radionuclide (accelerator produced).
Figure 2 The radioisotopes of iodine to the
right of the stable 127I are said to be neutron
rich and are mainly produced by reactor
irradiation. The isotopes to the left are said to
be neutron deficient and are mainly produced
by accelerators. However, there are many
exceptions. An accelerator may produce 131I
and 125I is in fact routinely produced by
reactors.
Reactors for radionuclide production
A nuclear reactor is a facility in which fuel such
as natural uranium or uranium enriched in 235U,
233
U, or 239Pu undergoes fission. In the process,
neutrons are produced from about 10 MeV and
downwards. The neutrons are moderated and in
different parts of the reactor one may find
different qualities of neutrons. The neutron has
no charge and does not feel the Coulomb
barrier. Even neutrons with very low energy
(thermal neutrons = 0.025 eV) can enter the
target nucleus and cause a nuclear reaction. On
the other hand, reactors do not produce very fast
neutrons (En < 12 MeV). Neutrons are not able
to knock out many nucleons from the nucleus
since the mean binding energy of nucleons is in
the range 8 MeV (see Chapter 2).
A reactor produces a neutron cloud in which we
place our target. Usually, we irradiate the target
isotropically. One has to consider that heat is
evolved in the reactor core and the temperature
at irradiation positions may easily reach 200 oC.
The reactor is characterized by the energy
spectrum, the flux (neutrons/cm2/s), and the
temperature at the irradiation position.
88
Figure 3 A top view of a nuclear reactor core
used for radionuclide production (TRIGA II).
Some of the tubes seen are part of a pneumatic
rabbit system for placing samples in different
positions of the core for irradiation. The targets
are encapsulated into closed tubes adopted for
the transport system.
Nuclear reactors especially designed for
radionuclide production and for neutron
activation are available. Some may even be
found in research hospitals for the clinical use
of short-lived radionuclides. Reactors used for
radionuclide production usually have a rabbit
system (mechanically or pneumatic) to enter
target into irradiation position and to take the
target out. In Table 1, possible nuclear reactions
for radionuclide production with a reactor are
reviewed.
The most typical neutron reaction is the
gamma capture i.e. a thermal neutron is
captured by the target nucleus and since there is
no momentum in the reaction the decay energy
is emitted as a prompt gamma ray. The reaction
in Table 1
59
Co (nth, γ) 60Co
is an important application when producing
strong γ-emitting sources for external therapy,
but is more or less of no value when labeling
biomolecules. Since the produced radionuclide
is of the same element as the target the specific
radioactivity, i.e. the radioactivity per mass of the production yield, one needs to work with
sample, is very low in this type of nuclear expensive enriched targets which, however, can
reaction. The ideal production route is when the be reused several times.
produced radionuclide is of a different element
than the target. This gives what is called a non- A general feature with reactor-produced
carrier added production route. radionuclides is that most emit high-energy β-
particles that give relatively high absorbed
Table 1 Typical nuclear reactions in a reactor doses to patients. However, there is a possibility
used for radionuclide production to produce neutron-rich radionuclides that do
not emit beta particles. An example is given
Type of Reaction T½ σ (mb) below
Neutrons
59
98
Mo(n, γ)99Mo (T½=66 h)
99mTc (T½= 6 h)
Thermal Co(nth, 5.3 a 2000
γ)60Co

14
N(nth, p)14C 5730 a 1.75
33
S(nth, p)33P 25 d 0.015
35
Cl(nth, p)35S 87 d 0.19

35
Cl(nth, α)32P 14 d 0.05

fission
131I 8d

24
Fast Mg(n, p)24Na 15 h 1.2

35
Cl(n, α)32P 14 d 6.1
The reaction 6Li(n, α)3H has a large cross-
section for the production of 3H+-ions of some
energy. These ions can be used for the
production of e.g. 16O(3H, n)18F. The target used
may be 6LiOH. In such a way, a reactor may
produce some typically neutron-deficient
radionuclides. A drawback with such a
production is that the crude reaction solution is
heavily contaminated with 3H-water that might
be difficult to remove.
Another reactor-produced neutron-deficient
radionuclide can be exemplified by the
production of 125I
124
Xe(nth, γ)125Xe (T½=17 h)
125I (T½= 60
d)
A drawback with this production is that 124Xe
has a natural abundance of 0.1 %. To increase
Figure 4 A cross-section of a 99Mo/99mTc -
generator (Courtesy of Mallinckrodt Medical
Inc.)
The 99Mo can also be produced by fission. The
system 99Mo/99mTc forms a generator system.
The mother nuclide 99Mo has a reasonably long
half-life for centralized production and
distribution to hospitals.
The generator system consists of a column on
which 99Mo is attached. By eluting the column
with saline the daughter nuclide 99mTc is
removed with negligible losses of 99Mo. The
generator system can work for one week before
it has to be renewed. By “milking” the generator
once a day, the hospital obtains supply of a
short-lived radionuclide which has ideal γ-
energy (140 keV) for the detectors, gives low
radiation dose to the patient (the decay of a
meta-stable nucleus gives mainly γ rays and few
conversion electrons).

89
 choose a nuclear reaction that is also practical
Inlet needle and economical.
for eluant
Elutions
Aluminium
crimp cap 1.0
99 Mo
Rubber stopper
0.5
Glass casing

Glass wool
0.2 99m Tc

99Mo
0.1
absorbed
onto aluminia
0.05
Glass filter

Rubber stopper 0.02

Aluminium
0.01
crimp cap Figure 7 A cyclotron that can accelerate
0 12 24 36 48
Outlet needle
hours
protons up to 17 MeV; generally used for
for 99mTcO4-
radionuclide production.

Figure 5 A cross- Figure 6 Elution profile
section through the of a 99Mo/99mTc-
generator column generator
Accelerators for radionuclide production
An accelerator can be huge as in CERN (with a
diameter of more than 1 km) and used for
particle physics. However, when using
accelerators for medically useful radionuclides,
much lower energy is needed. One can do very
well with a proton accelerator of < 80 MeV or
even less (< 40 MeV protons) if one also has the
possibility to accelerate deuteron- and alpha-
particles. During the last decade, small
accelerator yielding 16 MeV protons and 8
MeV deuterons have become fairly common at
larger hospitals for positron emission
tomography (PET) applications.
Although such accelerators are much compact
than a reactor, the particle energy they can
provide is 2-8 times higher than for the reactor
neutrons. That energy transferred to the nucleus
is much higher and more particles can be kicked
out of the nucleus. We may produce nuclides
farther from the stable nuclide line and we have
better opportunities to choose a suitable
radionuclide for our specific application. Since
we have more bombarding particles to choose
between, we also have a better opportunity to
Another advantage with accelerator production
is that, since we irradiate with protons or heavy
ions, we produce a compound nucleus that is of
a different element than the target. It is often
much easier to produce non-carrier added
products in an accelerator than in a reactor.
A technical difference between reactor and
accelerator irradiation is that in the reactor the
particles come from all directions but in the
accelerator there is a direction of the particles.
The number of charge particles is often smaller
and is usually measured as an electric current in
µA (6*1012 particles/s for protons but 3*1012 for
alpha-particles which have 2 charges per
particle).
A drawback in accelerator production is that the
charged particles are stopped much more
efficiently than neutrons. Protons of 16 MeV are
stopped in 0.6 mm Cu. If we have a typical
production beam current of 100 µA and a beam
area of 2 cm2 we stop 1.6 kW in a volume of 0.1
cm3 which will evaporate all types of material if
not efficiently cooled. The acceleration of the
beam is done in vacuum but the target
irradiation is made outside in atmospheric
pressure or in gas targets at 10-20 times over
pressure.

90
 He-gas Table 3 Cyclotron produced non- positron
emitting radionuclides. Examples from current
25 µm folie Solid target research program in Uppsala.
Particles from Water cooling
accelerator in of target
vacuum PET- T½ Decay Application
Nuclide Area
114
Cd(p, n)114mIn 50 d EC Therapy
197
He-gas Au(p,2p5n)191Pt 2.8 d EC Therapy
25 µm folie Gas target under pressure
α α-source
209
Bi(p,2n)208Po 2.9 a
Particles from
accelerator in 209
Bi(α,2n)211At 7.2 h α, EC Therapy
vacuum
Water cooling of
the target walls The accelerator production offers great
flexibility in producing one radionuclide in
Figure 8 Schematic targets for radionuclide different ways as seen in the example below,
production. The upper part of the figure which gives some of the available ways to
illustrates the use of solid target while the lower produce 123I (T½ = 13 h).
figure the use of a gas target. 127
I (p, 5n) 123Xe
123I
124
Table 2 Cyclotron produced positron-emitting Xe (p, np) 123Xe
123I
123
radionuclides. Examples from current research Te (p, n) 123I
122
program in Uppsala. Te (d, n) 123I
124
Te (p, 2n) 123I
121
Sb (αα, 2n) 123I
PET- T½ β+ Application 121
Sb (3He, n) 123I
nuclide (h) (%) area 123
Sb (3He, 3n) 123I
14
N(p,α)11C 0.34 100 Org. synthesis
18
O(p,n)18F 1.16 97 Halogenation
Factors to consider are economy, radionuclide
45
yields, amounts of radionuclide impurities,
Sc(p,n)45Ti 3.9 85 Chelators separation, and radiochemical aspects.
52 52
Cr(p,n) Mn 134 29 Mn kinetics
58
Ni(p,3p4n)52Fe 8.3 56 Fe kinetics Nuclear physical considerations
58
Ni(p,α)55Co 17.5 76 Chelators
If we want to observe an object, we usually
60 61
Ni(d,n) Cu 3.4 61 Cu kinetics illuminate it with light. The way the light is
76
Se(p,n)76Br 16.2 54 Halogenation reflected, diffracted and absorbed informs us
85
Rb(p,3n) Sr 83
32.4 24 Sr kinetics about the object and its properties. We may
88 86
learn whether it is transparent or opaque. If it is
Sr(p,3n) Y 14.7 33 Chelators transparent we can measure the refraction index,
110 110
Cd(p,n) In 1.15 62 Chelators and if it is opaque we can find its absorption
124 124
Te(p,n) I 100 23 Halogenation capacity. If monochromatic light is used, we
may find how these properties vary with
wavelength. The object may emit radiation of a
Usually, one uses thin foils (about 25 µm thick)
different kind (e.g. photo effect emitting
to separate vacuum and the target. Two such
electrons). If the light beam is shut down, the
foils are used with a stream of He-gas in-
object may continue to reemit light
between to cool the foils. He-gas is preferable
(phosphorescence).
since it has excellent cooling capacity and little
radioactivity is produced in the gas.

91
If we want to “see” an object as small as the
atomic nucleus, we need to use “light” of a
much higher frequency, which also means much
higher energy. However, the same type of
experimental approach can be used although we
need to design special particle detectors to
replace the eye. It is not by chance that one of
the most powerful nuclear models is called “the
optical model”. We can regard particle beams of
electrons, protons, neutrons and mesons as
“matter beams” that have wavelengths suitable
to “see” the atomic nucleus. These beams may
be reflected, refracted and absorbed too.
dΩ
dσ
IoN dΩ
dΩ
N
Φ such as (p,3n). Each such reaction further
Incident beam Io
Target Scattered
particles
Figure 9 A principle experimental setup. A
nuclear physicist is usually interested in the
particles coming out, their energy and angle
distribution, but the radiochemist is mainly
interested in the transformed nuclides in the
target.
A large difference between neutrons and charge
particles is their possibility to penetrate the
nucleus (Figure 10).
NEUTRON
CROSS-SECTION
PROTON
ENERGY
Figure 10 General cross-section behavior for
nuclear reactions as a function of the particle
energy. Since the proton has to overcome the
coulomb barrier there is a threshold that is not
present for the neutron. Even very low-energy
neutrons can penetrate into the nucleus to cause
a nuclear reaction.
92
We may write a nuclear reaction in the
following way
a+A
b+B+Q
where a is the incoming particle and A is the
target nucleus in their ground states (the
entrance channel). Depending on the energy and
the particle involved we may open different
outgoing channels, where b is the outgoing
particle and B is the rest nucleus in excited
states. Q is the reaction energy and can be both
negative and positive. If the incoming particle is
absorbed we have a capture process type (n,γ)
and in a reaction of type (p,n) we obtain charge
exchange. If many particles are expelled we can
refer to the reaction according to the process
distinguished by their excitation state is called a
reaction channel.
Different reaction mechanisms can use the same
reaction channel. Here we differentiate between
two ways
• The formation of a compound nucleus
• Direct reactions
The compound nucleus has a large probability
to be formed in a central hit of the nucleus. The
incoming particle is absorbed and an energy-
rich compound nucleus is formed. It has usually
a lifetime of about 10-14 seconds. This time is
long enough for the nucleus to forget the
direction of the incoming particle. However, the
nucleus needs to get rid of its energy excesses,
and sooner or later one or several particles
“evaporate” from the nucleus. The rest nucleus
formed is usually left in an excited state and
cools off further by emitting gamma-radiation.
The out-coming particles are emitted
isotropically since the compound nucleus has
forgotten about the direction of the incoming
particle. The energy of the emitted particles has
a typical distribution (evaporation spectrum).
Most emitted particles have a low energy but
some can have up to 5-6 MeV.
 Elastic reaction
(p, p)
Incident Compound
nucleus
proton
Direc
(p, p´) inelastic
t re a c
(p, d) picup
tions
(p, n) charge exchange
etc
Inelastic reactions
Figure 11 A schematic figure showing some
reaction channels at proton irradiation.
The direct reactions preferably occur at the edge
of the nucleus (see Figure 12). The incoming
energy is directly transferred to a nucleon
(knock-on reaction) giving two out-coming
particles or you may have a stripping reaction of
type (d,n). The outgoing particles usually have
high energy and are emitted in about the same
direction as the incoming particle.
a + A C
Before Delay
b the area of both the nucleus and the incoming
a + A B +
Before After
Before After
Figure 12 An illustration of the difference
between direct nuclear reactions (lower part of
the figure) and the formation of a compound
nucleus (upper part of the figure).
Most reactions are probably a mixture of these
two types of reactions. The probability of these
two events varies with energy in a different
way. The direct reactions are heavily associated
with the geometrical size of the nucleus. The
compound nucleus, however, is formed just
across the reaction threshold and the probability
is high when the particle energy is just above
(see Figure 13).
Threshold
56Fe
Cross section (barn)
(n, n´)
Total
Compound
Direct reactions
0 1 2 3 4 5 6 7 8
MeV
Figure 13 A schematic view of particle energy
variations of cross-section for direct nuclear
reactions and for forming of a compound
nucleus.
When producing radionuclides, a detailed
knowledge of different reaction probabilities is
a necessity. The reaction probability is
expressed as a cross-section or a surface. The
unit is barn (b), which is an area of 10-28 m2.
B This is a rather large unit, so mb is a more
Compound commonly used unit. The cross-section unit is
After nucleus related but not identical to the nucleus projected
formation area. The probability of a hit is a combination of
particle. Particles of low energy can act like a
b mass wave of low wavelength and cover a large
area in space. One may then obtain cross-
Direct section values of magnitudes higher than the
stripping area of the nucleus.
After hitting the nucleus, one has also to
consider the physical and statistical laws of
Direct distributing the energy so that a specific
knock-out reaction channel is opened. Some cross-sections
may therefore be very small. Figure 14
illustrates a well-known thumb rule in
radionuclide production. The maximum cross-
sections found at about 10, 30 and 40 MeV for
the (p,n)-, (p,3n)- and (p,4n)-reactions,
respectively. Thus, it takes about 10 MeV to
kick out a nucleon, i.e. a proton of 50 MeV can
cover radionuclide productions that involve the
emission of about 5 nucleons. Figure 14 also
shows an example of using the cross-section
information for an optimal production of 73Se
with protons. At low energy there is a disturbing
production of 75Se and if one uses excessively
93
high proton energy one obtains another
unwanted radionuclide impurity, 72Se. The last
impurity can be avoided completely by
restricting the proton energy to an energy lower
than the threshold for the (p, 4n)-reaction. The
impurity from the (p, n)-reaction is not possible
to avoid but can be minimized by using a target
thickness that avoids the lower proton energies
(having the highest (p, n)-cross-sections).
75As(p,n)75Se
103
75As(p,3n)73Se
102
101
75As(p,4n)72Se
100
0 10 20 30 40 50
Proton energy, Ep (MeV)
Figure 14 Excitation functions of
75
As(p,xn)72,73,75Se reactions. The optimal
energy range for the production of 73Se is Ep =
40
30 MeV.
As seen in Figure 14, the chosen production
parameters are a compromise. A proton range
from 40
30 MeV uses the (p,3n) cross-
section well. Some 72Se-contamination is
acceptable in order to increase the yield of 73Se.
An important factor is the half-life, 75Se
(T½=120 d), 73Se (T½= 7.1 h) and 72Se (T½=
8.5d). Sometimes it is possible to wait for the
decay of the radioactive contaminants. This is
not the case here, instead the longer half-lives of
the radioactive contaminants help to keep the
radioactivity of these low. The cross-sections
give a number of the produced atom nuclei of
each isotope. If the half-life is long, then the
decay is spread out over a longer time, hence
the radioactivity is lower than for a short-lived
radionuclide. The relation between the number
of nucleus produced and radioactivity is set by
the specific radioactivity.
The practical setup when doing a radionuclide
production looks like this. A suitable As-target
is made and irradiated with 40 MeV protons.
The thickness of the target is such that it
decreases the proton energy down to 30 MeV.
94
This then gives a radioactivity yield of the
desired radionuclide at end of bombardment
(EOB) mainly dependent on the beam current
and the irradiation time. The yield is usually
expressed in GBq/µAh, i.e. the produced
radioactivity per time integrated beam current.
If possible, one tries to keep the radioactivity of
the contaminants small (< 1 %). However, after
EOB, the long-lived radio-contaminants start to
increase with the decay of the product.
Specific radioactivity
In a target irradiation, we produce a number or a
mass of radioactive atoms. The most common
way to express this number is to give a measure
of the radioactivity in Bq. However, this is an
indirect way since it only tells us how many
nuclei decay per second. To tell how many
radioactive nuclei we had from the start, we
need to add up all decays from the start to
infinity.
Radioactive decay is a statistical process. If we
have a number of radioactive nuclei N they all
have the same probability of decaying. This
probability does not change in time, nor is it
affected by chemistry, environment etc. During
a small time, dt, the number of decayed atoms is
dN. The radioactivity of the sample is expressed
as A = dN/dt. The relative number decayed,
λ∗dt = dN/N is always the same and is constant.
λ is called the decay constant and has the unit
seconds-1. We can write this as a differential
equation (the minus sign is due to the decrease
of N in the decay process).
dN/dt = - λ * N
This equation has the unique solution
N = No * e-λ*t
where No is the initial number of radioactive
nucleus.
Since the radioactivity A = dN/dt we get a
relation between radioactivity and the number
of radioactive nucleus, A = λ * N. This means
that the radioactivity also will decrease in the the labeling chemistry does not differ between
same way as the number of radioactive atoms e.g. the radioactive 131I and the stable 127I, we
have to include the number of stable iodine
A = Ao * e-λ*t atoms that might have been in the target or
added during the separation process.
where Ao is the initial radioactivity at time 0.
Spec. Radioact. = Ao/(No + Nstable)
In the decay process, the half-life (T½) of the
radionuclide, is an important parameter. Its is Since in many cases the stable atoms are more
defined as numerous than the radioactive, the mass does
not change with time but the radioactivity does.
A = Ao 2-t/T½ The specific radioactivity decreases with time.

The relation between λ and T½ is given by In radionuclide production and radiochemical

separations the following expressions are used:
λ = ln(2)/T½ Carrier free: Only the radioactive atoms are
present in the preparation, i.e. no stable isotopes
By integrating the radioactivity decay curve of the same element are present.
(Figure 15, left) from t= 0
∞, we add
together all the decayed nuclei, No = ∫A dt No carrier added: In the separation process, no
(Figure 15, right). stable isotope of the same element has
deliberately been added. However, one cannot
EXPONENTIAL DECAY DECAY INTEGRAL
exclude that some stable istope may have been
RADIOACTIVITY FRACTION

1 1
DECAYED ATOM FRACTION

0,8 0,8
added through uncontrolled processes, such as
stable carbon being added from glass wear etc.
0,6 0,6

0,4 0,4
Carrier added: In the separation process one
0,2 0,2 deliberately adds some amounts of the same
0 0
element to facilitate or stabilize the separation
0 1 2 3 4 5
0 1 2 3
TIME IN T½
4 5
TIME IN T½ process. This will decrease the specific
radioactivity of the product..
Figure 15 By integration of the exponential
decay curve (left figure) one obtains a number For radioactive labeled compounds one usually
of totally decayed atoms (right figure). calculates specific radioactivity as radioactivity
per number of labeled and unlabeled molecules.
The relation between radioactivity and the This means that we might here obtain values
number of atoms that are producing this higher than the theoretical values for radioactive
radioactivity is called specific radioactivity atoms since we may have more than one
radioactive label per molecule (i.e. from 14C-
Ao ln(2) labeled benzene where all six atoms are 14C-
Spec. ac t. = =
No T ½ atoms).
The number of radioactive atoms can be To illustrate how the theoretical specific
expressed in mass units such as gram or moles, radioactivity is calculated, the following
and specific radioactivity can then be expressed example is given.
in units such as MBq/g or GBq/µmole.

If we only consider the radioactive atoms, we

can always give a unique value of the specific
radioactivity. If we include the stable isotopes
we have to modify the concept somewhat. Since

95
 radionuclides of other elements from our
Example. How to calculate the maximal specific preparation.
radioactivity of 125I.
Figure 16 A hot-lab
The physical half-life of 125I is T½=59.41 days. equipped with a hot-
The number of radioactive atoms is then cell for high
59.4*24*3600/ln(2)= 7.41*106atoms/Bq. radioactivity work.
We consider the radioactivity 1 GBq = 109 Behind 5-10 cm
disintegration/s. This correspond to 7 .41*1015 lead one can work
atoms. By using the Avogadro´s number we get with a radioactivity
that 1 µmol of 125I-atoms corresponds to of 100 GBq.
6.023*1017 atoms. We can then calculate the Automated systems
specific radioactivity (SA) of 125I or manipulators are
used to protect
6.023 * 1017 personnel.
SA(125I)= = 81 GBq/µmol
7 .41 * 1015
Time is essential. If we work with a short-lived
In literature, specific radioactivity is often given radionuclide such as 110In (T½ = 69 minutes)
in the old unit for radioactivity, Curie (1 Ci=3.7 and the separation procedure takes 4-5 hours,
1010 Bq). the radionuclide is of no practical interest. The
separation procedure should take at most about
1 Ci = 3.7 1010* 59.4 * 24 * 3600 / ln(2) = 1 half-life.
2.74*1017 atoms
C15O2 C15O
17
6.023 * 10 Deutron-irradiation
SA(125I)= = 2.2 Ci/µmol
2.74 * 1017 TARGET Fast on-line
d+14N-->15O+n chemistry
Radiochemistry
15O
2 H215O
During target bombardment, a heavy shielding N2+1% O2
is necessary around the reactor/accelerator to
protect from prompt radiation. But even after Figure 17 Target system for production of 15O
shut down, radiation shielding is advisable. (T½= 2 minutes) and on-line production of a
Besides the wanted radioactivity, the target may number of labeled ligands.
contain several other reactions, either of the
same element (radio-contaminants) or other Gaseous or liquid targets are disadvantageous in
elements. Some radionuclides may be short- the irradiation but advantageous in the
lived and before processing it may be of separation since the separation system may
advantage to let them decay. In the subsequent easily be automated.
processing, the target is usually transported in a
lead shield to a hot-laboratory and placed in a
1 2
lead shielded hot-box.
4 5
In the radiochemical process we want to remove 3
the small amount of desired radioactivity (≈ 1100ºC 200ºC
nanomole) from the bulk of target (0.1-1 g). We 6
want the separation efficiency to be as high as Argon
possible and the separation time as short as
possible. We also want to separate all Figure 18 A schematic description of the 76Br
separation equipment. (1) Furnace, (2)

96
Auxiliary furnace, (3) Irradiated target, (4)
Deposition area of selenium, (5) Deposition Table 4 Specific radioactivity of some
area of 76Br, (6) Gas trap. commonly used radionuclides in biology and
nuclear medicine
Solid targets usually have to be dissolved
(usually in boiling acids) before wet chemical Radio- T½ Specific radioactivity
separation methods are applied. In general, two Nuclide Common values
principles are used: liquid extraction and ion Max for compounds
exchange. Occasionally, thermal separation GBq/µmole GBq/µmol
techniques may be applied, which have the 3
H 12 a 1.08 0.01-5
advantage that they do not destroy the target 14
(important when expensive enriched targets are C 5730 a 0.0023 0.0001-0.01
35
used) and that they lend themselves to S 87 d 55.5 0.0001-0.01
automation. 32
P 14 d 340 0.001-10
33
P 25 d 197 0.001-1
As an example of such dry methods the thermal 131
separation of 76Br (T½ = 16 h) is described. The I 8d 599 0.01-1
target is Cu276Se, a selenium compound that can 125
I 60 d 81 0.01-1
withstand some heat. The nuclear reaction used 57
Co 270 d 18 0.1-10
is 76Se(p,n)76Br. 58
Co 71 d 68 0.1-10
75
1. The target is placed in a tube and heated, Se 120 d 40 0.001-0.1
under a stream of argon gas, to evaporate 197
Hg 64 h 1776 0.01-1
the 76Br activity by dry distillation (Figure 203
Hg 47 d 104 0.001-1
18). 11
C 20 min 3.55 *105 1-100
18
2. A temperature gradient is applied to F 2h 6.59 *104 1-100
separate the deposition areas of 76Br and
traces of co-evaporated selenide in the tube
by thermal chromatography.

3. The 76Br activity deposited on the tube wall

is dissolved in small amounts of buffer or
water.

Separation yields of 60-70% are achieved and

the separation time is about one hour. Since dry
distillation permits the extraction of
radiobromine without destroying the target, the
Cu2Se targets are reusable. Considering the
rather expensive 76Se-enriched target material,
this is practically a prerequisite for this type of
production. The chemical form of the 76Br
activity after separation, analyzed by ion-
exchange high-performance liquid
chromatography (HPLC) and thin-layer
chromatography (TLC), was almost exclusively
found to be bromide.

97
 CHAPTER 10
NUCLEAR BIOLOGY AND MEDICINE: IMAGING
Imaging techniques have always been important
in the application of radionuclides and tracer
techniques. Henri Becquerel did not only
discover the natural radioactivity but he also, by
chance, invented the autoradiographic technique
when he discovered the impact of the uranium
salts on a photographic film. This technology
has developed considerably and can today give
quantitative information of tracer distributions
from the organ level to the cell nucleus
structures. External detection of gamma-
emitting nuclides has developed from scanning
with a collimated gamma sensitive probe to to-
day’s sophisticated systems for Single Photon
Emission Computed Tomography (SPECT) and
Positron Emission Tomography (PET).
Autoradiography
Autoradiography in biomedical applications is a
technique for the visualization of radioactive
materials in cells, cell cultures, tissues and
organs. A thin sample (cell culture) or a thin
section obtained by tomographic techniques is
placed in close contact with a detector that can
register the position of the decays.
Sample thick
thin
Detector thin
thick
Eββ high Eββ high Eββ low
Sample thick Sample thin Resolution good
Resolution bad Resolution good independent of
independent if if film is thin sample and film
film thick or thin thickness
Figure 1 General principles of autoradio-
graphy. The sample and the detector in close
contact give the best resolution and sensitivity.
If spatial resolution is desired in the same
range as e.g. the beta particle range, then the
thickness of sample and detector is of less
importance. If the desired resolution is better
than the range of emitted particles, then the
thickness of sample and detector are the
dominating factors. An important factor is of
course the intrinsic resolution of the detector.
98
Several factors, such as the thickness of sample
and detector as well as the energy and
penetration ability of emitted radiation,
determine the information one can obtain from
such a system (Figure 1). Other important
factors are how firm the radioactivity is bound
to the sample. For example, a labeled ligand
that diffuses in the sample during preparation
gives a low-resolution image.
Several sample preparation techniques for
different applications are possible to use, such
as
• Macro-autoradiography (Whole Body). The
animal is frozen in a liquid that prevents
freeze artifacts. A frozen block is cut by a
saw or placed in a whole body tomograph.
Sections with a thickness of 20 µm or more
are mounted directly on a plastic tape.
• Embedded sample, tomographic section.
Normal histological embedding and
mounting techniques on glass are used.
Tomographic sectioning gives samples with
a thickness of 3 µm or less.
• Frozen sample, tomographic section, freeze-
dried mounting on glass backing. This
technique can be used when the labeled
ligands are expected to diffuse. Samples can
be obtained with a thickness of about 3-5
µm.
• Ultra-tomographic techniques used in
electron microscopy. Sample thickness of
about 100 nm.
The thinner the sample, the smaller the
radioactivity contents in the sample. One has to
adopt the radioactivity concentration in the
sample to the desired resolution. When using
ultra-thin sections, one also has to consider the
specific radioactivity of the ligand and whether
it is theoretically possible to obtain detectable
radioactivity amounts in the sample.
Generally any radionuclide can be used for
autoradiography. However, if high resolution is
important, then one prefers to use radionuclides
that emit radiation with low penetrating ability
such as 3H and 125I. These radionuclides can be (about 100 nm). Ideally a monolayer of silver
used to label a variety of biomolecules at high bromide crystals is used and exposed. After the
specific radioactivity. development procedure the silver grains are
hardly visible even in light microscopy with the
Photographic film best resolution. However, in electron
microscopy the high atom number of silver
Photographic emulsion is still the dominant gives a good contrast and images of a high
detector. It is also the only choice when spatial resolution may be obtained.
resolution of a few µm or less is desired. There
is a large choice of different types from standard
X-ray films to liquid film emulsions for high
resolution. Measuring the film density can be
used to quantify the autoradiography, e.g. by a
computerized image analyzer system. A
calibration curve of (radioactivity
concentration)*(exposure time) to density can
then be used. It is important to obtain a correct
exposure of the film due to the limited linear
range of the system.

B Figure 2 Whole-body
autoradiography
showing the distribution
T of 14C-methionine.
L u Some structures are
L i marked, such as brain
(B), lungs (Lu), liver
(Li), intestine (I) and Figure 3 Electron microscope autoradiogram
tumor (T). showing labeled acetylcholine on receptors in a
The density can be neuromuscular junction. More information at
T measured and related to www.nbb.cornell.edu/neurobio/salpeter/salpeter.html
I radioactivity
concentration by
comparing with a Phosphorous imaging techniques
calibration scale.
The photographic film has some disadvantages
such as

• Low sensitivity, which means long exposure

If closer contact is needed the film emulsion
without backing can be applied directly onto the
sample. The two techniques used here are the
strip film technique and the dipping technique.
Another way to quantify is to measure the
number of silver grains in the film emulsion. It
is often necessary to do manually using a
microscope and it is a tedious work. An
alternative is to use an electron microscope. A
diluted fine-grain film emulsion is used. Using
the surface tension in a loop, a thin film is
produced that is applied onto a thin sample
times.
• Low linearity. The exposure has to be right.
If the film is over- or underexposed, one
needs to redo the process. Sometimes it is
difficult to cover the dynamics in the sample
by one single exposure.
• Time consuming handling. The samples
have to be dry before applying the film
emulsion. The developing process takes
time. Digitalization or grain counting is
necessary in order to quantify.

99
Lately, new types of detectors have been
developed to overcome such drawbacks. They
are based on different physical principles such
as multi-wire chamber (gas detector), multi-
channel detectors (scintillator detector), and
phosphorus imager (trapped excited electrons
similar to thermo-luminescence detectors), strip
detectors and pixel detectors (solid state
detectors). Their resolutions are limited to about
25 µm and upwards. Here, we will just describe
one type of these detectors, the phosphorous
detector, in some detail. Figure 5 The read-out as a function of number
of disintegration for a phosphorous system and
An image plate of this type contains photographic film.
BaFBr:Eu+2 crystals. When a radioactive sample
is placed on the plate the emitted ionizing
radiation excites electrons in the crystal. In the
de-excitation process, a small fraction of
electrons are trapped in an excited stage and a
latent image of the radionuclide distribution is
formed.

Scanning
Laser (red)
Unstable state

Trap
PM-tube
Ground state
Figure 4 Principle of the phosphorus imaging
technique.
A scanning red laser beam is used to “develop”
the imaging plate. The size of the laser beam
spot determines the resolution (25-200 µm).
The energy in the red light photons is enough to
lift up the trapped electrons to the unstable
state. It has then a high probability to fall down
to the ground state emitting a blue light photon
that is counted by a photo-multiplier tube (PM-
tube). The number of blue photons registered in
each position is proportional to the radioactivity
concentration of the sample. Exposing the
image plate to an intensive white light will
anneals remaining trapped electrons and the
image plate can be used again.
Figure 6 The read-out system of one of the
commercial systems for phosphorous imaging.
A red laser spot is scanned over an exposed
image plate by an optical system.
The technique is more sensitive than the film
since a denser material is used as detector. It is
also more useful for high-energy particle for the
same reason. Besides, it is linear over a larger
range of exposures than the film.
Gamma camera
The Anger gamma camera is used to detect
single photons. A main component is the lead
collimator (Figure 7) which selects the photons
that are emitted perpendicular to a large
NaI(Tl)-crystal.

100

Figure 7 A schematic view of the gamma
camera.
Photons of another direction are ideally
absorbed by the lead in the collimator. A photon
hitting the crystal may give rise to scintillation
(a number of light photons). This light spark is
viewed by a number of PM-tubes. Depending
on their position they “see” varying numbers of
light photons. Knowing the position of the PM-
tubes we can calculate the x, y-coordinates of
the scintillation and the energy deposited in the
crystal.
Each photon detected by the crystal can then be
registered, and an image of the radionuclide
distribution can be constructed from a number
of such events. The technique lends itself to
dynamic investigations.
The weak point in the gamma camera is its
collimator. It is made up of a slice of lead (3-10
cm thick dependent on the desired performance)
with thousands of narrow holes. From an
isotropic source the collimator picks out a very
small fraction of photons (the exact number
depending on the thickness and the hole
dimensions). This means that the counting
efficiency for the system is low.
The intrinsic resolution of the gamma camera,
e.g. how well the system can decide the position
of the scintillation, is about 4 mm. However, if
one made a collimator with the same resolution,
101
its sensitivity would be unacceptably low.
Typical values are in the order of 15 mm at a
distance of 10 cm from the collimator. The
resolution varies with the depth.
Salivary glands
Liver
Bladder
Front Back
Figure 8 Diagnosis of Disseminated Malignant
Pheochromocytoma using 123I-Meta-
Iodobenzyl-Guanidine (MIBG).
The collimator also sets restrictions to the
gamma energy, which one may use. High-
energy photons may penetrate the thin lead
walls between the holes (the septa) of a high-
resolution collimator. For high gamma energies,
the septa have to be bigger and the resolution of
the collimator decreases. High-energy photons
may also penetrate the crystal without giving a
scintillation, which decreases the sensitivity.
Optimal gamma-ray energies to be used in
typical gamma camera systems are in-between
80-200 keV. One of the most used radionuclides
is 99mTc (140 keV).
The gamma camera image is a depth integrated
radionuclide distribution. Gamma rays from
different depth are detected with different
sensitivity and resolution. Gamma rays from
different depth are also absorbed and scattered
in different ways. Hence, the gamma camera
gives information that is difficult to quantify in
absolute terms.
SPECT
Each gamma camera exposure gives a depth-
integrated measurement of the radionuclide
distribution. In Figure 8, the patient is viewed
from two sides: the frontal projection and the
back projection.
Figure 9 The gamma camera is stepwise
rotated around the patient (typically 3.6 o per
step). For each step a projection is obtained,
i.e. a measurement of the integrated
radionuclide distribution from that angle.
If we have a time stable radionuclide
distribution, i.e., we have reach an equilibrium,
the two measurements should give about the
same radioactivity distribution, however
mirrored (Figure 9). The difference is due to
somewhat different attenuation of photons.
By a proper mathematical treatment of all these
measurements (about 50 projections) it is
possible to describe the radionuclide
distribution in 3D. We obtain what is called
tomographic information since we slice the
patient, not by knife but with the help of the
computer. For each slice we can present the
radionuclide distribution as an image. The most
common reconstruction method is called
filtered back-projection.
102
Figure 10 Perfusion measured in a normal
female volunteer using SPECT and 99mTc-
HMPAO. Data are presented in 20 tomographic
slices through the brain.
Figure 11 A dedicated SPECT system. Three
gamma camera detectors rotate together in to
speed up the data collection.
A typical time to make a SPECT investigation
is about one hour. The time per projection is
about one minute, and 50 projections or more
are needed. The long exposure time also
requires that the radionuclide distribution is
“frozen”, i.e. does not change substantially
during the investigation time. To speed it up
two or more gamma camera heads can be placed
together as in Figure 11.
PET
In positron emission tomography (PET)
positron-emitting nuclides, such as 11C are used.
The positron travels 2-3 mm in tissue before it
is stopped. When stopped it attracts an electron
and forms a strange atom, a positronium, which In PET, data is collected in all directions
exists for a short while before annihilation of randomly. There is no need to move the detector
the electron masses. The energy is emitted as but all projections are collected simultaneously.
two anti-parallel 511 keV photons as indicated Each event is sorted into a set of parallel
in Figure 12. coincidence channels defining one projection.
This means that a full set of data for
Positron Decay tomographic information may be collected
Coincidence Detection during 2-10 seconds only. There is no need for a
15N
radionuclide distribution in steady state but very
15O tube fast kinetic investigations can be performed.
r|PM
ν V Ph
oton detecto
5 11 ke
positron β+
TRANSMISSION
hoton
or 11 keV P
e |detect 5 Rotating
P M tu
b
e - electron source
Coincidence Detection
Time Window: 10 ns (BGO-detectors)

Figure 12 The emission of two annihilation

photons in the positron decay.

detector rings

Bed

Detectors

annihilation photons Figure 14 A transmission scan. A line source of

Figure 13 A schematic view of a positron
camera system.
The photon energy of 511 keV is high. It comes
out of the patient efficiently but it penetrates
lead and detector materials almost equally well
too. It is difficult to collimate these photons as it
is done in the gamma camera, and NaI(Tl) is not
efficient enough. In PET, one or several rings of
small detectors are placed around the patient
(Figure 13). Each one of these detectors is
coupled in coincidence with all opposite
detectors. If two such detectors are hit
simultaneously (within 10 ns) the PET camera
assumes that it has been hit by two photons
originating from the same decay. A straight line
is defined by the known positions of the two
detectors and the PET camera assumes that the
decay has occurred somewhere close to that
straight line.
68
Ge/68Ga is circulated around the patient. The
density of a patient slice is seen viewing lungs,
heart, spine and muscle. The bed upon which
the patient lies is of a light material with low
density and is hardly seen.
At present, the most common detector material
is bismuth germinate (BGO), a scintillating
crystal with a high Z-number (Z=83) and a
density close to 8 g/cm3. These properties make
a detector with a fairly high efficiency to 511
keV photons. A crystal with the length of 25-30
mm has a detection efficiency of about 70 %,
which in the coincidence channel corresponds
to 50 % efficiency of detecting coincidences.
There is no need for a lead collimator. Instead,
we can say that we use an “electronic
collimator” by measuring the two opposite
directed photons by coincidence. This makes
the system rather sensitive
Commercial systems with a spatial resolution of
about 4 mm in all directions are available today.
The resolution is mainly set by the crystal

103
dimensions but there are factors, such as the attenuation in the body and can then obtain a
range of the positrons that set the theoretical correct quantification within a few per cent.
limit of resolution to 2-3 mm.

An advantage of PET is that the radionuclide

distribution can be quantified in absolute values
(Bq/cm3). In an investigation, two
measurements are made: transmission and
emission. The transmission is performed as
described in Figure 14.

EMISSION

Figure 15 An emission scan of a human heart.

A ligand (acetate) labeled with a positron
emitting nuclide (11C) is given.

A line source containing a long-lived positron

emitter is rotated around the patient. We still
use the coincidence technique. One of the
photons hits a detector directly whereas the
other has to pass through the patient before it
hits an opposite detector. We call this a
transmission scan since we have an external
source and one of the photons is transmitted
through the patient. Since we view the patient
from all angles we have the same situation as in
an X-ray investigation using computerized
tomography (CT). We measure the density of
the patient but with the same photon energy as
we do later on in the emission scan.

The line source is removed and the ligand

labeled with a positron emitter is injected. Both
photons have to pass through parts of the
patients and we call this an emission scan. By
combining the transmission scan and the
emission scan we are able to correct for

104
 CHAPTER 11
Labeling methods
A correct labeling of biomolecule is important
for the outcome of all experimental applications
of nuclide technique. The measuring technique
is just giving information about the radioactivity
but the labeling technique couples this
information to the information we want – the
fate of the biomolecule.
Important general aspects of labeling have already
been mentioned in Chapter 9. Here we will go into
particulars about labeling of biomolecules.
Protein labeling through tyrosine
The most common labeling method is to label
tyrosine with halogen isotopes. The tyrosine can
either be labeled as a part of a protein (direct
labeling) or as a part of a precursor, which is then
attached to the protein in a suitable reaction
(indirect labeling). Halogens such as iodine and
bromine are introduced into the ring structure of
tyrosine by oxidative processes. Iodine might also
be attached to tryptophan or sulphydryl groups. At
a high pH (>8) histidine might also be labeled.
The most commonly used agents for the oxidation
is Chloramine-T either in solution or immobilized
on polystyrene beads (Iodogen). This method is
used for labeling with radioactive isotopes of
iodine such as 125I for basic biology and 131I for
medical applications such as imaging and nuclide
therapy. The last years, 123I for planar gamma
camera/SPECT as well as 124I for PET, have also
been applied using the tyrosine labeling strategy.
Chloramine-T is the sodium salt of N-monochloro-
p-toluene-sulfonamide (see Figure 2). It is a mild
oxidizing agent. In an aqueous solution it releases
hypochlorous acid, which oxidizes iodine probably
to an active iodonium ion [H2OI+]. This ion is
incorporated into the tyrosine residue of the
proteins. The exact mechanism for this reaction
with carrier-free radioactive iodine isotopes is not
known in detail.
Enzymatic labeling
Enzymes such as lacto-, chloro- or bromo-
peroxidase can be used to label protein. One
example is the labeling of antibodies with
105
radioactive isotopes of bromine through the use
of bromoperoxidase. The reaction is initiated by
the addition of H2O2. After a suitable reaction
time, the reaction is ended by addition of
sodium metabisulphite. A drawback is that it is
often difficult to purify large protein product
from the enzyme. This is usually easy in the
tyrosine labeling method described in Figure 2
since only low-molecular-weight reagents are
used.
Figure 2 Labeling of tyrosine with iodine by the
Chloramine-T method.
a) The structure formula for Chloramine-T.
a) CH3 SO2 N Cl Na+
O H Cloramine-T O H
and 125I-iodide 125I
b)
Oxidation of
the iodide
gives H2O125I+
CH2 CH2
R1 -NH - CH - C - R2 R1 -NH - CH - C - R2
O O
b) A schematic view of the labeling procedure.
The oxidative reaction is, after a suitable
time stopped by adding sodium bisulfide.
Indirect protein labeling
Amino group labeling
The so-called Bolton-Hunter reagent provides a
general method to label amino group. This
reagent labels proteins at the epsilon amino
groups (i.e. lysine) and can, but more slowly,
also react with the alpha amino group at the
amino terminus of a protein or polypeptide. The
Bolton-Hunter reagent, already labeled, is
commercially available. The reagent contains a
tyrosine ring that carries the iodine and a
succinimidyl group that can react with amino
groups (Figure 3).

Figure 3 The Bolton-Hunter reagent is seen in the
upper part to the right. It is pre-labeled in the ring
with radioactive iodine. The succinimidyl group
reacts with the amino group on the lysine and
thereby an indirect labeling is achieved.
Several Bolton-Hunter "like" reagents have been
described. All contain the succinimidyl group
while the tyrosine ring is modified to allow for
different properties. Thereby one can counteract, at
least partly, degradation or dehalogenation in vivo.
Figure 4 The reagent is pre-labeled with 3H
through a carbon chain. The succinimidyl group
reacts with the amino group on lysine. The protein
is "tritiated".
It is also possible to modify the ring so that not
only iodine but also the halogens bromine and
astatine can be coupled. Furthermore, there are
similar reagents where the ring is exchanged for
other groups and which allow for 35S or 3H
labeling. It is still a succinimidyl group that
accounts for the binding to the amino group of
the protein (Figure 4).
106
Thiol labeling
If the succinimidyl group in the above-
mentioned reagents is exchanged for a
maleimide, the reagents can be used for reaction
with thiol groups. In principle, this means that
reactions can be made with free SH groups in
the protein (i.e. cystein). Such reagents can
contain for example 3H or 14C and are
commercially available.
Conjugations
There are numerous biochemical conjugation
methods described in the literature.
Conjugation, in this context, generally means to
bring molecular components together. Thus, any
method by which two components (e.g.
carbohydrate to protein, protein to protein,
protein to nucleic acid etc) are coupled together
can be used as a labeling method if one of the
components is pre-labeled. One example of
conjugation mediated labeling is that
radiolabeled carbohydrates are connected to
amino groups of proteins via activation of the
carbohydrate (using e.g. activation agents such
as periodate or CDAP). Several methods to use
COOH and NH2 groups in conjugation
chemistry have been described and all such
methods are of potential interest for
radiolabeling.
Chelators
The coupling of chelators to antibodies or other
proteins and the subsequent addition of
radioactive metals that are trapped by the
chelator is one possibility for protein labeling.
The requirement is that suitable radioactive
metals are available.
Many types of chelators are described in the
literature and the most often used is DTPA
(diethylene triamine penta-acetic acid) which
binds metals such as indium (110In for PET,
111In for SPECT) and yttrium (89Y or 90Y,
both for nuclide therapy). DTPA is also, for
some applications, coupled to ligands such as
the eight-amino acid somatostatin analogue,
octreotide, giving the commercially available
product OctreoScan suitable for characterization harvested, homogenized and the protein purified
of tumors with somatostatin receptors. from the homogenate. The advantage of
biosynthetic labeling is that no extra chemistry
Chelator has to be applied and the product protein does
DTPA not suffer from conjugation or other labeling-
induced modifications. The disadvantage is that
Protein
it is often difficult to obtain a high specific
Conjugation radioactivity. Radioactive amino acids can be
chemistry purchased commercially containing, for
example, radioactive carbon (14C) or hydrogen
(3H).

Protein Add radioactive

111In Protein aminoacid
Chelation

Figure 5 Labeling of a protein with 111In by

the use of the chelator DTPA.
Synthesis
There are many different types of chelators
(DTPA "like" and others) with different
substituted side groups to allow for coupling via
different biochemical methods. The advantage
of using chelators is that all chemistry involved
can be made a long time in advance whereas the
addition of the radioactive metal can be made
directly before the use of the whole complex Harvest
(Figure 5). A simple "desalting" is then enough radioactive protein
to get rid of the excess radioactive nuclides.

Biosynthetic protein labeling

Figure 1 Biosynthetic labeling. Radioactive
amino acids are added (top), the protein
Most labeling methods modify the molecule to
synthesis produce labeled proteins (middle) and
some extent, which may change the molecules
the product are harvested for purification
behavior regarding e.g. distribution and binding
(bottom).
properties. A way to evade this and to label
exact copies of biomolecules (e.g. amino acids,
Labeling of nucleic acids
anti-bodies, structural proteins, and enzymes)
with radioactive nuclides is to feed a cell culture
Nucleic acids can be labeled with several
with amino acids containing radioactive
different methods. The rapid development in the
nuclides. The requirement is of course that the
area of DNA techniques have speeded up the
cultured cells synthesize the proteins of interest.
development of both "radioactivity" and
The radioactive precursors are given to the cell
"fluorescence" based methods for labeling and,
culture medium with the cells. If the cells
in some cases, interesting combinations of these
excrete the protein, the culture medium can be
approaches. The main radiolabeling methods
harvested (Figure 1) and the product purified.
used are nick translation labeling, 5´ end
The cell culture can be used repeatedly. If the
labeling, 3´ end labeling, hybridization labeling
protein is not excreted, the cells have to be
and biosynthetic labeling. Furthermore, special

107
methods to label small primers or
oligonucleotides (e.g. random primer labeling)
exist. The field is still developing and new
methods might be expected in the near future.
Nick-labeling
Nick labeling means that intact DNA, in a cell-
free system, is exposed to enzymes, which
opens up the double stranded DNA. Short
segments of single stranded DNA (only 5-8
bases) are formed. These single strand DNA
segments are restored to double strand DNA by
enzymatic "repair synthesis" using e.g.
radiolabeled thymidine in this process. Thus, the
DNA is labeled with radioactivity but is not
chemically modified since only enzymes from
the normal repair processes are used.
5´ end-labeling
5´ end labeling means cell-free labeling of the 5´
end of DNA or an oligonucleotide often using
polynucleotide kinase to add phosphate at the
end. The radioactive nuclides in the added
phosphate can be 32P or 33P (high- and low-
energy beta emitters, respectively).
3´ end labeling
3´ end labeling is a method that uses e.g.
deoxynucelotidyl transferase to add radioactive
dATP to the end of DNA or an oligonucleotide.
dATP is, in these applications, labeled with 32P.
There are also special reagents (e.g. ddATP) that
ensure that only one labeled nucleotide is added
to the end, and there are special types of dAPT
that are labeled with 35S instead of 32P.
Hybridization based labeling
As described above, oligonucleotides can be
labeled with nick translation or end labeling. If
such oligonucleotides are made single stranded
they can then be used for hybridization to RNA
or to partially or totally denatured DNA if there
are homologous sequences. This is a powerful
technique to find and label certain genes (base
sequences). However, the procedure leaves the
hybridized nucleic acid blocked in certain
sequences by the oligonucleotide probe and this
108
probably disturbs the function if the DNA is
used for further tests. Thus, this type of labeling
is convenient for analytical purposes if certain
base sequences are to be found, but it might not
be a good labeling method if the nucleic acid
must be functional afterwards.
Biosynthetic labeling
A direct way to label nucleic acids, DNA or
RNA, is to feed a cell culture with radioactive
thymidine or uridine, respectively. The
radioactive precursors are given to the cell
culture medium and the cells are allowed to
grow in physiological conditions. After a
suitable time (often 1-24 hours) the cells are
harvested, homogenized and the nucleic acids
purified from the homogenate. The advantage of
such labeling is that no extra chemistry has to be
applied and the product does not suffer from any
chemical modifications. The disadvantage is
that it is difficult to obtain high specific
radioactivity. The radioactive precursor is
usually 14C- or 3H-labeled.
The nucleic acids can also be labeled with 32P-
phosphate, using a biosynthetic route but in this
case many other cellular components are also
labeled by general phosphorylation processes.
Special labeling methods
Labeling of arginine
Arginine residues in proteins have a general role
in positively charged recognition sites for
anionic ligands. This is especially the case when
the ligands contain important carboxyl- or
phosphate groups. Arginine also occurs in the
active site of many enzymes, especially those
enzymes that catalyze phosphorylation. Arginine
can be labeled with radioactive phenylglyoxal,
which reacts with the guadino group of arginine.
It is claimed that proteins after such treatment
are more resistant to trypsine-mediated
cleavage.
Carbohydrate labeling (OH)
Carbohydrates with free OH groups can be
labeled via enzymatic or oxidizing
transformation of the OH groups to aldehyde
groups, which then can react with radiolabeled
sodium-, potassium- or cyano-borohydride.
Another alternative is to allow the aldehyde
groups to react with radiolabeled aniline
forming a Schiffs base, which is then stabilized
with unlabelled borohydride. Radioactive
reagents for such reactions are commercially
available.
Fast labeling with 11C, 15O and 18F for PET
applications
Very fast methods have been developed for
labeling of low-molecular-weight sub-stances
with 11C (T1/2=20 min) (e.g. methionine), 15O
(T1/2=2 min) (e.g. O2) and 18F (T1/2=110 min)
(e.g. fluorodeoxy-glucose). The short physical
half-lives of these nuclides make it necessary to
apply fast organic chemistry processes, and
several such methods have been developed by
organic chemists working with PET
applications. The nuclides mentioned here are
mainly used for diagnostic purposes in PET
based applications.
Labeling with "long lived" positron emitters.
There are several so-called "long lived" positron
emitters, such as 55Co (T1/2=18 hours), 76Br
(T1/2=16 hours) and 124I (T1/2=4 days). They
are suitable for labeling of macromolecules with
long biological turnover rate like antibodies.
Using such positron emitting labels and PET
will give a detailed pharmaco-kinetic
information of such molecules, which is of
importance in diagnostics, therapy and in the
development of new drugs. The physical half-
lives of these nuclides are sufficiently long that
standard protein labeling methods can be
applied. Such methods are chelator-based
labeling for the metal ion 55Co and for the
halogens, 76Br and 124I, direct or indirect or
enzymatic protein labeling can be applied. The
use of such nuclides has so far been limited but
with improved possibilities for macromolecular-
based gene-, immuno- and nuclide-therapy, the
requirements to follow macromolecular
distributions with PET cameras are being met.
109
Labeling of dextran
Dextran and other polysaccharides can be
labeled via activation with periodate or CDAP
(to activate the OH groups) so that any amine-
containing radiolabeled substance attaches. This
has, for example, been applied in the attachment
of radiolabeled glycine or tyrosine to dextran.
Another labeling method is based on the fact
that dextran in nature is synthesized by the
enzymatic addition of glucose from the
disaccharide sucrose. The sucrose is cleaved by
the enzyme sucrase, which then adds the glucose
part on to the 6´ end (the growing end) of
dextran. The fructose part of sucrose is left in
the monomer form. Thereby dextran slowly
grows to form large chains. By introducing
radiolabeled sucrose, which must be labeled on
the glucose part, radioactive glucose attaches to
the dextran. This gives a "normal" not
chemically modified dextran that is
radiolabeled. These methods are of interest since
dextran in some cases will probably be used as a
part of conjugates in tumor therapy for targeted
treatments and thereby will serve as a carrier for
drugs and nuclides (Figure 6).
EGF-receptors
10B dextran
131
I 111
In
76
Br
211
At
EGF-dextran can
be labeled with
EGF Tumour cell
various nuclides
Figure 6 Schematic picture of radiolabeled
EGF-dextran binding to the EGF-receptor
(EGF=epidermal growth factor).
Purification
After radiolabeling, the product must be
purified. Usually this is a simple step but
sometime it might be necessary to consider
various sophisticated purification methods.
The purification is in most cases only a question
of removing excess radionuclides and the
reagents applied. In many cases the product is a
macromolecule whereas the nuclides and
reagents are of low molecular weight. In that
case, it is often enough to use a small, simple
"desalting" column to purify the product and at
the same time, if necessary, change the buffer
system. In other cases it might be more
complicated, such as after enzymatic labeling or
when the product is of low molecular weight. It
might also be complicated to purify if the
labeling technique induces heterogeneity; e.g.
some product proteins are labeled whereas
others are not or when other forms of the
product are produced due to the labeling
chemistry.
When a simple desalting technique is not
adequate, one has to find more sophisticated
methods. A number of preparative
chromatographical methods are available such
as large-sized exclusion columns with good
molecular weight separation properties, ion
exchanger columns, affinity or hydrophobic
interaction columns or other methods such as
preparative electrophoresis or precipitation
techniques.
Quality control
After purification or storage of a radiolabeled
product, it is often necessary to perform quality
control to ensure that the product has the same
properties as a standard. All available analytical
methods in chemistry and biochemistry can then
be considered as well as functional biological
test.
Storage conditions
Radiolabeled products might be destroyed due
to oxidation or radiolysis. Oxidation damage is
most common for sulfur-containing substances
such as methionine and cystein. Storing the
compound in nitrogen atmosphere (e.g. in liquid
nitrogen) can minimize oxidation. Radiolysis
means that the substance irradiates itself and
thereby causes its own decomposition.
There are several ways to reduce radiolysis. The
compound may be stored diluted. A common
110
way is to store the compound in a refrigerator
or, if it can stand it, frozen. Some
macromolecules are in fact very sensitive to
freezing and a large fraction might be
biologically inactivated. Long-term storage can
also be made in the frozen state using
cryoprotective agents (DMSO, sucrose, and
glucose), which minimize the risk of ice crystal
formation. However, the general rule is to use
fresh preparations and to avoid long-term
storage.
Stability during incubations
Another problem to consider is that radioactive
substances might be modified during biological
experiments. The substance might, in the
experimental situation, be exposed to reduction,
oxidation or enzymatic degradation. Such
processes might be of interest to study in the
experiment. However, such processes might
also be undesired, and destroy or make the
planned study difficult to interpret. The stability
of radioactive substances can, in principle, be
controlled through the extraction of biological
material during the experiment. The samples are
then analyzed for the chemical form of the
radioactivity to allow for analysis of degradation
products.
Labeling services
If possible one should buy your radiolabelled
chemicals commercially instead of doing the
labeling yourself. There are several commercial
companies that offer a large collection of
especially low molecular molecules like amino
acids and nucleotides. When working with
macromolecules it may be more cost effective to
buy kits for labeling and then do the labeling in
your own laboratory. In house labeling is also
necessary e.g., when the applied nuclides have a
very short physical half-life (as in many PET
applications). Another case is when a new
substance are discovered and synthesized in
your laboratory. The pioneering laboratory must
then analyze and characterize the labeled
molecule e.g. to investigate how the labeling
procedure affects the biological function and
behavior. This is a research topic in itself.
 CHAPTER 12
Applications
Accuracy and sensitivity
Radioactive nuclides give the possibility to
study biological and medical processes with
high accuracy and sensitivity. The accuracy can
be very high since the measurements are, in
most cases, not only quantitative but also
possible to calibrate in detail against standard
preparations, and this can be made for very low
concentrations of substances. The sensitivity is
very high since single decays can be detected.
This allows the presence of only a few labeled
molecules (and even single molecules) to be
detected. The applications given below are
divided in two groups: biological applications
(basic biology and preclinical investigations
using to a large extent pure beta emitters) and
diagnostic nuclear medicine (applying gamma
and positron emitters).
BIOLOGICAL APPLICATIONS
DNA synthesis
DNA synthesis in cells or tissues can be
quantitatively studied through the use of
radiolabeled thymidine (Figure 1).
O
X RNA synthesis
N X CH3 (3H)
CH3 (14C)
O by cells for the synthesis of all forms of RNA.
N 125I, 76Br,
dR and other
(deoxyribose) halogens
Figure 1 Radiolabeled thymidine and iodinated
and brominated analogs
Thymidine easily passes the cell membrane. It
is, in the cell, only used for the synthesis of
DNA and for no other macromolecular
structure. Thus, it provides a very specific way
to detect which cells or cell populations have an
ongoing DNA synthesis. Scientific history is full
of exciting discoveries obtained with the help of
radioactive thymidine (characterization of the
cell cycle, analysis of the action of growth
111
factors and oncogenes etc.). Thymidine is
available as a commercial product labeled with
3H or 14C and those products are among the
most sold radiochemicals.
Genes
The presence of certain gene sequences in DNA
can be accurately analyzed by hybridization
experiments in which radiolabeled
oligonucleotides, containing the gene sequence
to be analyzed, are allowed to hybridize with
denaturated DNA. The presence of the
interesting gene can then be analyzed
quantitatively by measurement of the
radioactivity. Such analysis is called "southern
blot" when the reaction is made on a blotting
paper to which the DNA has been transferred.
The analysis is called in situ hybridization when
the DNA is still in the cell (or at least in a
chromosomal conformation).
Oligonucleotides for such analysis are most
often synthesized in a non-radioactive form in a
"DNA synthesizer" and are then labeled with
radioactive nucleosides by a process called nick-
or end labeling. In cell analysis, it is necessary
to first degrade RNA with RNAse to eliminate
the otherwise dominating binding to mRNA.
RNA synthesis can be studied by applying
radioactive uridine, which is specifically used
Uridine easily passes the cellular membrane and
the incorporation process is, principally, parallel
to the use of thymidine for studies of DNA
synthesis. Radioactive forms of uridine are
available commercially. Studies using uridine
give an overall measure of the total RNA
synthesis in the cell. For gene-specific analysis
of mRNA synthesis, see below.
Gene activity and transcription
The activity of a gene in terms of "transcription
activity" (synthesis of mRNA) can be measured
quantitatively through hybridization between a
radiolabeled oligonucleotide, containing the
gene sequence of interest, and the formed
mRNA with that gene sequence. Such analysis
is called "northern blot" when the reaction is
made on a blotting paper to which the mRNA
has been transferred and in situ hybridization
when the mRNA is still in the cell. Undesired
binding to DNA is normally not a problem as
long as the DNA is in double stranded form. If
necessary, DNA can be degraded with DNAse.
Oligonucleotides for such analysis are made the
same way as described above for gene analysis.
Specific proteins
The presence of specific proteins or other forms
of biological material in cells can be analyzed
using radiolabeled antibodies with specificity
for the protein to be analyzed (see also below
regarding sandwich techniques with labeled
secondary antibodies). The biological sample is
exposed to the radioactive antibody and when
the protein is present the quantity can be
determined through analysis of the radioactivity.
Such analysis is called "western blot" when the
reaction is made on a blotting paper to which
the biological sample has been transferred. It is
called radioimmuno detection when the analysis
is made directly on a more or less intact
biological sample. Many forms of radioimmuno
detection are described in the chapter on
antibodies. Protein specific antibodies can often
be purchased from companies specialized on
immunology products. In most cases, it is
necessary to do the radioactive labeling at one’s
own laboratory using methods as described in
Chapter 9. In a few cases, it is possible to buy
already labeled antibodies.
Protein synthesis
The overall protein synthesis in cells or tissues
can be studied accurately through the use of
radioactive amino acids. The amino acids are
incorporated in the proteins and thereby give an
intracellular accumulation of radioactivity. All
most common types of amino acids are
commercially available in radioactive forms.
It should be noted that different amino acids are
taken up with different efficiency over cell
membranes but this is normally not a big
problem in basic biology studies. Cocktails of
112
different amino acids are sometimes used for the
measurement of total protein synthesis. If the
synthesis of only specified types of proteins is
studied, then the use of radioactive amino acids
must be combined with other methods, such as
gel electrophoresis or immunoreactions.
Phosphorylation and basic metabolism
The regulation of energy flow in biological
systems involves phosphorylation and de-
phosphorylation (AMP ADP ATP). The energy
flow is to a large extent related to energy
generation in the glucolysis and the citrate cycle,
and the energy consumption to the synthesis of
different molecules and the active transport of
molecules. In all cases, it is possible to study the
metabolic steps by using 32P or 33P labeled
phosphate, PO4. The radioactive phosphate is
given to cells and tissues. After different
incubation times the compounds of interested
are isolated with biochemical methods (column
chromatography, thin layer chromatography,
electrophoresis, etc) and their content of
radioactivity is measured. . The appearance and
disappearance of radioactivity reflects
phosphorylation and dephosphorylation,
respectively. The analysis is easily quantified so
that the number of molecules involved in the
processes can be estimated. Radioactive
phosphate is commercially available at
reasonably low prices and is one of the most
used radioactive chemicals.
Phosphorylation and signaling
One of the most interesting processes studied in
recent years is phosphorylation related to
intracellular signaling. Many intracellular signal
systems seem to be mediated via
phosphorylation and dephosphorylations. One
example is receptor-ligand interactions (e.g.
growth factor-receptor), which initiate primary
phosphorylation of the receptor itself and related
proteins leading to cascades of phosphorylation
to exert signal transduction, often all the way
into the cell nucleus. In the last steps, the
phosphorylation initiates DNA mRNA
transcription (activation of genes). Such
phosphorylation can easily be analyzed by using
32P labeled phosphate. The radioactive
phosphate is given to cells that exert the studied
reaction. Thereafter, the studied molecules are
isolated (most often using electrophoresis) and
the amounts of 32P associated to the molecules
are a measure of the phosphorylation. By
incubating the cells for longer times with
radiolabeled phosphate, dephosphorylation
processes can also be studied.
Phosphorylation in cell free systems
Radioactive phosphate can be used for end- or
nick-labeling of nucleic acids in cell-free
systems. This allows detection of the DNA
when processed with different biochemical
methods for purification, size determination and
sequencing. Radioactive phosphate is, as
mentioned above, commercially available and at
reasonably low prices so there are no severe
economic limitations for the use of the
described methods.
Glucolysis and energy metabolism
The amount of glucolysis can be measured
through the use of radioactive glucose.
However, "natural" glucose is quickly
metabolized and does not give an accumulation
in cells and tissues with high glucolysis. Instead,
the synthetic analog deoxyglucose is used since
it is to a large extent phosphorylated in cells and
might bind irreversibly to key enzymes in the
glycolytic pathway and thereby accumulate and
allow accurate detection. Radioactive
deoxyglucose without phosphorylation easily
passes cell membranes and is commercially
available. This is the most often used precursor
when energy metabolism is analyzed. The steps
in the "citrate cycle" (or Krebs cycle) can be
analyzed through the use of other radioactive
tracers.
Glycosylation
The glycosylation of any macromolecule in a
biological system can be analyzed by adding
radioactive carbohydrates of the suitable type
and then analyzing how much radioactivity is
associated with the studied macromolecule. One
limitation is the accuracy by which the studied
113
molecule can be isolated through available
purification methods.
Other metabolic processes
Carbohydrate and lipid synthesis can be studied
by using more or less specific radioactive
precursors. Methylation processes can be
studied by applying relevant radiolabeled
precursors and melanin synthesis can be studied
through the use of radiolabeled thioureas. Pre-
formed melanin can be analyzed with several
substances since melanin is used by the body as
a scavenger for toxic agents. Radiolabeled
analogues of such agents might then be used for
the analysis. The list of possibilities for
metabolic studies using radioactive substances
can be made very long and it seems that most
metabolic processes can be studied if action is
taken to radiolabel relevant precursors. In
principle, at least one precursor must be known
and radiolabeled and this precursor then serves
as a starting point for the studies under
consideration. The limitations thereafter are the
limitations of the biochemical methods to
analyze metabolites of the radioactive test
substances. An economic limitation might be
the high costs to develop a radiolabeling
technique for a new substance.
Uptake and processing of new compounds
In compound development (e.g. development of
new chemotherapy agents, metabolic inhibitors
or targeting agents for radionuclide therapy) it is
necessary to characterize the biological
properties of the compounds in some detail.
This means analysis of the cellular binding,
internalization, retention and degradation as
well as the in vivo properties such as half-life in
the systemic circulation, uptake and metabolism
in the liver, uptake and release from the kidneys,
and uptake in many other organs. By labeling
the compound with radioactive nuclides, it is
possible to follow such processes. If the
compound is of low molecular weight, the most
attractive approach is to exchange stable
nuclides, most often carbon or hydrogen, for the
same type of nuclides in the radioactive form, in
this case e.g. 14C or 3H. With this procedure
the chemical and biological properties of the
compounds are not disturbed. If the compound
is macro-molecular (protein, glycoprotein,
carbohydrate etc.) it is often possible to label
with, for example, radioactive iodine without
disturbing the biological properties of the
molecule too much. The availability of
radiochemical reagents for the radiolabeling
varies from case to case.
Receptor-ligand interactions
If the binding of a ligand (e.g. a growth factor)
to a receptor is characterized in some detail it is
convenient to use a radioactive ligand. The
binding of the radioactive ligand can be
quantified in detail and related to the number of
cells or the mass of the biological sample under
physiological conditions. The specificity of the
radiolabeled ligand can first be analyzed using
native non-radioactive ligand in excess (at least
tenfold excess) during the time of analysis. If
the radiolabeled ligand binds specifically, the
binding should under such conditions be
displaced more or less completely. If there are
unspecific inter-actions between the radioactive
ligand and the biological test material, the signal
of the radioactivity cannot be efficiently
displaced. In cases when the radioactive ligand
binds specifically, one can, at 4oC, analyze
binding versus concentration of the added ligand
(to obtain saturation) and also gradually displace
the radioactive ligand by increasing amounts of
non-radioactive ligand. Such analysis gives
input data for a so-called "Scatchard analysis" in
which the affinity and the number of binding
sites can be quantitatively determined for the
ligand-receptor interaction. Such applications
are of importance in the development of
targeting agents for imaging and/or therapy of
spread tumor cells and metastatic growth.
Antibody-antigen interactions
If the antibody binding to an antigen is
characterized in some detail it is, in the same
way as described above for ligand-receptor
interactions, convenient to use a radioactive
antibody. The specificity of the radiolabeled
antibody can be analyzed using native non-
radioactive antibody in excess. If the
radiolabeled antibody binds specifically, the
114
binding should under such conditions be
displaced more or less completely. As for
ligand-receptor interactions one can also in this
case analyze binding versus concentration, at
4oC, and gradually displace the radioactive
antibody by increasing amounts of non-
radioactive antibody to obtain input data for
"Scatchard analysis". An application is, also in
this case, the development of targeting agents
for imaging and/or therapy of spread tumor cells
and metastatic growth.
DIAGNOSTIC IN NUCLEAR MEDICINE
Development
From the historical point of view, the initial and
most common strategy for the diagnostic use of
radionuclides has been to give e.g. 99mTc (pure
gamma emitter) in salt form (or associated to
some biomolecule) and then to study the uptake
pattern in normal healthy persons. The spatial
distribution of the radioactivity was studied in
pictures obtained using different types of
gamma cameras. Different diseases could then
be studied by analyzing aberrations in relation to
the so-called "normal" pictures. One striking
example is the studies of the breakdown of the
blood brain barrier. It was found that this barrier
was locally disrupted in patients with malignant
gliomas, and analysis of this is today an
established type of investigation.
Today, the use of radioactive nuclides for
diagnosis of different diseases is very advanced.
Radioactive precursors can be synthesized for
specific studies (as described above in the
section on biological applications) and the
spread of these precursors can be followed more
or less quantitatively over the body by using
advanced scintigraphy (e.g. SPECT and PET).
The spatial resolution is so good (in the order of
1 cm) that one can fairly well calculate the
uptake of radioactivity in different tissues and
organs. The metabolic processing of the
precursors can sometimes be followed by
chromatographic analysis of blood and urine,
and counting of the amount of radioactivity in
the different molecular preparations.
Basic biology versus clinical tests
In principle, the same radioactive precursors as
described above for basic biology studies can
also be applied in clinical patient studies to try
to analyze aberrant processing in the studied
metabolic steps. The main difference is due to
the fact that in the clinical work, gamma
radiation has to be used to allow for external
detection with high efficiency and precision.
Thus, the pure beta emitters (3H, 14C, 32P and
35S) often used in basic biology are of no or
limited interest since it is in most cases not
possible to obtain tissue samples for analysis.
Instead, high-energy gamma or positron (giving
rise to gamma radiation after annihilation)
emitters have to be applied. Examples of such
nuclides are 11C, 15O, 18F, 76Br, 99mTc,
110In, 111In, 123I, 124I and 131I. The fact that
some of these nuclides have a very short
physical half-life puts special requirements on
the radiolabeling and the handling of the
nuclides. It also gives certain advantages since
few radioactive molecules need to be attached to
the test compound to have a certain number of
radioactive decays during the time of analysis.
The discussion on clinical applications below
follows, due to pedagogical reasons, the same
order as for the biological applications. This is a
somewhat unusual order in comparison to what
is used in clinical textbooks on nuclear medicine
where simpler methods such as the use of free
iodine for studies of uptake in the thyroid are
mentioned first and more complicated
investigations employing macromolecules are
mentioned later. However, to start with
discussions on genes and DNA-mRNA
transcription (as in most biological textbooks) it
is clearly seen how much more biologically
advanced all laboratory techniques are in
relation to the available clinical methods. This is
hoped to serve, for the reader, as an inspiration
for further efforts to develop the clinical
methods.
DNA synthesis can probably be fairly well
studied in different tissues in patients through
the use of 76Br labeled thymidine (11C labeled
thymidine is probably less suited because of the
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short half-life of 11C). The technique is fairly
new, and tests are in progress to analyze DNA
synthesis in growing tumors. Brominated
thymidine (Figure 1) has to be synthesized at
special laboratories designed for fast synthesis
of radioactive nuclides.
Genes and transcription
The presence of certain gene sequences in DNA
and the corresponding DNA-mRNA
transcription activity may, in the future, be
analyzed in the intact body via hybridization
techniques. Oligonucleotides containing the
gene sequence to be analyzed should then be
labeled with radioactivity and injected into the
patient. The analysis at the clinic should in most
cases be non-invasive and made by external
monitoring of the localization of the applied
radiolabeled oligonucleotides. Of course, there
are still many unsolved problems relating to this
problem like: How can the oligonucleotides find
their way to the target organs? How can they
find their way over the cell membranes to finally
reach the nucleic acid of the targeted cell? How
to prevent degradation of the oligonucleotides
before they reach the target? The expected
developments in gene therapy will hopefully
help to obtain suitable transport means for such
oligonucleotides. Thus, the technique is not
available today but will hopefully be available
in the future. The only realistic possibility
available today is the analysis of operation
material and biopsies with the same techniques
as applied in basic biology (in vitro diagnosis).
RNA synthesis
RNA synthesis can in principle be studied using
radioactive uridine in parallel to the use of
radioactive thymidine for studies of DNA
synthesis. However, the use of gamma emitters
for studies of uridine is not so developed, and
we therefore have to wait for further progress
from the involved research laboratories.
Specific proteins
Radiolabeled antibodies with specificity for the
protein to be analyzed can in some cases be used
to externally visualize the presence of the
protein in different parts of the body. Whether
the analysis can be made in a quantitative
manner depends on the availability of the
protein in the tissue, the antibody penetration
properties and on the scintigraphy technique
applied. This technique, often called
immunoscintigraphy, radioimmunolocalization
(RIL) or radioimmunodiagnosis (RID), is
presently under intense development and we
foresee interesting applications in the near
future.
Protein synthesis
The overall protein synthesis in tissues and
organs can, at the clinic, be studied rather
accurately through the use of radioactive amino
acids. One amino acid used for PET-based
analysis is 11C labeled methionine which has,
for example, been used to monitor the increased
amino acid uptake (and possibly also increased
protein synthesis) in different types of tumors.
In principle, all types of amino acids can be
labeled with gamma or positron emitters and
applied in such studies. An interesting aspect is
to try to distinguish between amino acid
accumulation due to changed transport in certain
diseases and the changed protein synthesis. The
parallel use of amino acid analogs, one that can
only be taken up intracellularly and one that can
also be incorporated in proteins, might give
future possibilities to study this.
Phosphorylation
Phosphorylation processes in patients will
hopefully be studied in the future applying
phosphate analogs. This is at present the subject
for research and development.
Glucolysis and energy metabolism
The amount of glucolysis in different tissues can
be measured quite accurately applying 18F-
labeled deoxyglucose, often called FDG. The
uptake in different tissues is assumed to reflect
the amount of glucolysis, and it has in many
cases been found that tumors have an increased
uptake. Suitable precursors for studies of the
citrate cycle will hopefully be developed in the
future.
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Glycosylation and other metabolic processes
The glycosylation of any macromolecule can, in
principle, be analyzed by adding relevant
radioactive carbohydrate and then analyzing
how much radioactivity is associated with the
studied macromolecule. The limitation is, if
blood or urine samples are not enough for the
analysis, that tissue samples have to be taken
from the patient and the studied molecule
isolated through available purification methods
(chromatography, gel electrophoresis).
Carbohydrate, lipid and melanin synthesis and
methylation processes can be studied by using
more or less specific radioactive precursors and
following similar strategies as indicated for the
studies of glycolysation. Some progress has
been made along these lines but there is a lot of
developmental work before even a minor part of
all potential possibilities can be exploited.
Therapeutical compounds
In the development of new chemotherapy
agents, metabolic inhibitors and targeting agents
for radionuclide therapy, it is necessary to
characterize the biological properties of the
compounds. This is partly made in the
laboratory but it is also important to study
parameters like uptake in different tissues to
allow for estimation of the half-life in the
systemic circulation, uptake and metabolism in
the liver, uptake and release from the kidneys,
and uptake in many other organs. By labeling of
the compound with gamma- or positron-
emitting nuclides it is, in principle, possible to
follow such processes in the patient or in
volunteers. The investigations should be
complemented by chromatographic analysis of
blood and urine to give knowledge about the
presence of degradation products.
Molecular interactions
If the binding of a ligand or an antibody to a
receptor or antigen is to be characterized
directly in a patient, the difficulties are much
larger than when analyzing such phenomena in
the laboratory. The difficulties partly relate to
the large volume of the patient requiring
enormous amounts of compounds for studies of
saturation and displacement. It is also very
difficult to assure that the transport systems in
the body allow for interactions to take place in a
representative and reproducible way.
Furthermore, there are difficulties due to
metabolism of the labeled ligands and
antibodies (a correct "Scatchard analysis"
normally requires inhibition of degradation at
4oC). It is of course not possible to distinguish
degradation products from the original
substance through the necessarily non-invasive
analysis systems (gamma cameras). In spite of
these difficulties, scientists are seriously
attempting to develop models through which
scintigraphic dynamic data can be treated in
terms of molecular flow and interactions.

Analysis of operation material and biopsies

If, at the clinic, there are possibilities to obtain

fresh living tissue samples that either can be
cultured or prepared otherwise as any biological
sample in experimental biology, then all the
possibilities for analysis listed above (see
biological applications) are available. This is
often called in vitro diagnostics and is a growing
research area. However, we foresee an
interesting development regarding in vitro
diagnostics in the future.

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