Fluorescence

• luminescence, mostly found as an optical

phenomenon in cold bodies, in which the molecular
absorption of a photon triggers the emission of
another photon with a longer wavelength.
• The energy difference between the absorbed and
emitted photons ends up as molecular vibrations or
heat.
• Usually the absorbed photon is in the ultraviolet
range, and the emitted light is in the visible range,
but this depends on the absorbance of the
particular fluorophore
 Fluorescence
• Fluorescence is named after the mineral
fluorite, composed of calcium fluoride,
which often exhibits this phenomenon.

Fluorescence induced by exposure to ultraviolet light in vials containing

various-sized cadmium selenide (CdSe) quantum dots
Fluorescence occurs when a molecule or quantum dot relaxes
to its ground state after being electronically excited

Excitation:

Fluorescence (emission):
 Fluorescence quantum yield

• The fluorescence quantum yield gives the

efficiency of the fluorescence process. It is
defined as the ratio of the number of
photons emitted to the number of photons
absorbed
 Fluorescence quantum yield
• The maximum fluorescence quantum yield is 1.0
(100%); every photon absorbed results in a
photon emitted. Compounds with quantum yields
of 0.10 are still considered quite fluorescent
• Fluorescence quantum yield are measured by
comparison to a standard with known quantum
yield. The quinine salt, quinine sulphate, in a
sulphuric acid solution is a common fluorescence
standard.
 Fluorescence lifetime
• The fluorescence lifetime refers to the time the
molecule stays in its excited state before
emitting a photon. Fluorescence typically
follows first-order kinetics

• [S1]: remaining concentration of excited state molecules at

time
• [S1]0: initial concentration after excitation
This is an instance of exponential decay. The
lifetime is related to the rates of excited state
decay as:

∑ i ki is the sum of all rates of excited state decay.

Note: : A quantity is said to be subject to exponential decay

if it decreases at a rate proportional to its value.
Applications of
fluorescence
 Lighting
• E.g. common fluorescent tube
• Inside the glass tube is a partial vacuum and a small
amount of mercury.
• An electric discharge in the tube causes the mercury
atoms to emit light. The emitted light is in the
ultraviolet (UV) range and is invisible
• the tube is lined with a coating of a fluorescent
material, called the phosphor, which absorbs the
ultraviolet and re-emits visible light
 Organic oils
• Crude oil fluoresces in a range of colours,
from dull brown for heavy oils and tars
through to bright yellowish and bluish
white for very light oils and condensates.
• This phenomenon is used in oil
exploration drilling to identify very small
amounts of oil in drill cuttings and core
sample.
 Biochemistry & medicine
• Large biological molecules can have a fluorescent chemical group
attached by a chemical reaction, and the fluorescence of the attached
tag enables very sensitive detection of the molecule.
• is used in monitoring expression levels for thousands of genes
simultaneously in many areas of biology and medicine, such as
studying treatments, disease, and developmental stages.
• For example, DNA microarray can be used to identify disease genes
by comparing gene expression in diseased and normal cells.
• It is also used to assess large genomic rearrangements.
• DNA microarray is applied when looking for single nucleotide
polymorphism in the genome of populations.
• It is also used to determine protein binding site occupancy throughout
the genome.
Many biological molecules have an intrinsic
fluorescence that can sometimes be used without
the need to attach a chemical tag.
Sometimes this intrinsic fluorescence changes
when the molecule is in a specific environment, so
the distribution or binding of the molecule can be
measured.
Bilirubin,is highly fluorescent when bound to a
specific site on serum albumin.
Zinc protoporphyrin, formed in developing red blood
cells instead of haemoglobin when iron is
unavailable or lead is present, has a bright
fluorescence and can be used to detect these
problems.
 Fluorescence & plants
• Chlorophyll makes plants green, but it fluoresces a blood red
colour when exposed to ultraviolet light
• Natural fluorescent plants contain pigments like betaxanthis
which absorb shorter wavelengths of light exciting electrons to
a higher energy state and emit longer wavelengths of light as
the electrons return to the ground state.
• Plants have also been genetically modified to fluoresce.
• Fluorescing transgenes are used in plants as reporters of gene
expression in vivo.
• Fluorescent antibodies are used to visualize protein localization
in vitro.
 Fluorescence & plants

• Transgenic plants are being sent to Mars to which

reporter genes have been added allowing it to
glow for different environmental “stressors”.
• These stressors include temperature, drought,
disease, metal content in the soil, peroxides, etc.
• Each stressor will glow at a different wavelength
that will be monitored.
Jellyfish

Jellyfish contains some proteins which are intensely fluorescent on exposure to

ultraviolet light. They have been used to image calcium flow in cells in real time.
Green Fluorescent Protein (GFP) and related proteins are used as reporters for a
number of biological events

Some Minerals and Gems

Fluorescent rocks include fluorite, calcite, gypsum, ruby, talc, opal, agate, quartz,
and amber. Minerals and gemstones are most commonly made fluorescent or
phosphorescent due to the presence of impurities. The Hope Diamond, which is
blue, phosphoresces red for several seconds after exposure to shortwave
ultraviolet light.
• Club Soda or Tonic Water

The bitter flavoring of tonic water is due to the presence of quinine, which
glows blue-white when placed under a black light.

• Body Fluids

Many body fluids contain fluorescent molecules. Forensic scientists use

ultraviolet lights at crime scenes to find blood, urine, or semen which are all
fluorescent.

• Vitamins

Vitamin A and the B vitamins thiamine, niacin, and riboflavin are strongly
fluorescent.

• Antifreeze

Manufacturers purposely include fluorescent additives in antifreeze fluid so

that black lights can be used to find antifreeze splashes to help
investigators reconstruct automobile accident scenes.
Laundry Detergents

Some of the whiteners in detergent work by making clothing a bit fluorescent. Even
though clothing is rinsed after washing, residues on white clothing cause it to glow
bluish-white under a black light. Blueing agents and softening agents often contain
fluorescent dyes, too. The presence of these molecules sometimes causes white
clothing to appear blue in photographs.

Tooth Whiteners

Whiteners and some enamel contain compounds that glow blue to keep teeth from
appearing yellow.

Postage Stamps

Stamps are printed with inks that contain fluorescent dyes.

The cathode ray oscilloscope

The Cathode Ray Oscilloscope has a fluorescent screen which employs zinc silicate
as phosphor, emitting green light when struck by the electron beam
Phosphorescence
• Phosphorescence is a specific type of
photoluminescence related to
fluorescence.
• Unlike fluorescence, a phosphorescent
material does not immediately re-emit the
radiation it absorbs
• Phosphorescence is a process in which energy
absorbed by a substance is released relatively
slowly in the form of light.
• This is in some cases the mechanism used for
"glow-in-the-dark" materials which are "charged" by
exposure to light.
• Unlike the relatively swift reactions in a common
fluorescent tube, phosphorescent materials used
for these materials absorb the energy and "store" it
for a longer time as the subatomic reactions
required to re-emit the light occur less often.
S0: ground state of the fluorophore

S1: first (electronically) excited state

T1: triplet

Phosphorescent powder under

visible light, ultraviolet light, and
total darkness.
• Most examples of "glow-in-the-dark" materials do
not glow because they are phosphorescent.
• For example, "glow sticks" glow due to a
chemiluminescent process which is commonly
mistaken for phosphorescence.
• In chemiluminescence, an excited state is created
via a chemical reaction. The excited state will then
transfer to a "dye" molecule, also known as a
(sensitizer, or fluorophor), and subsequently
fluoresce back to the ground state.
• E.g of phosphorescent objects are toys, athletic
balls,etc.

Glow-in-Dark-Sticker
Thank you for your attention