International

ELF exposure systems for School of Biolelectromagnetics

in vitro and in vivo experiments “Alessandro Chiabrera”
1st COURSE “Methodology in bioelectromagnetic experimental investigations”
21 – 28 April 2004, Erice

ELF exposure systems

for in vitro and in vivo experiments

G.A. Lovisolo, L. Ardoino

Section of Toxycology and Biomedical Sciences

CR ENEA Casaccia, Roma
ELF exposure systems for in vitro and in vivo experiments

Summary

 General requirements of ELF exposure systems

 How to generate a uniform magnetic field?
 Systems of coils
 Basic concepts and definitions
 How to determine optimal values for the electric parameters
 First steps of improvements: most widely used “reference” systems
 Experimental and technical improvements
 Two examples of ELF exposure systems for in vivo and in vitro studies
ELF exposure systems for in vitro and in vivo experiments

Before listing the requirements, a simple consideration about ELF field

characteristic must be done…

At ELF the electric and magnetic part of EM field can be considered

acting in a separate manner.
An external electric field is greatly attenuated inside the body and
perpendicularly oriented to the surface. This is due to the dielectric
properties (conductivity and permittivity) of the body tissues.
On the contrary, the magnetic field penetrate the body virtually
unperturbed and induced electric fields and currents inside the tissues.

“the main objective of the bioeffects studies of ELF fields is

to investigate the effects related to the exposure to the
magnetic field, thus the exposure system has to be essentially
a system for generating magnetic fields”
 ELF exposure systems for in vitro and in vivo experiments

Basic requirements - all exposure systems -

 (Real time) Measurements of the generated fields and of the background

levels

 Shielding from external disturbances

 Allow sham exposure

 Dosimetry: assessment of induced electric fields and currents inside the

target (especially for in vitro experiments)

 Assessment and monitoring of environmental parameters and other

changes introduced by the system (electrical heating effects, noise and/or
vibrations produced by the coils, ...)
ELF exposure systems for in vitro and in vivo experiments

Basic requirements - ELF exposure systems -

 Modify intensity and frequency values of magnetic field generated in a

wide range (0 – 100 Hz).

 Large volumes of uniform magnetic field, related to the size of the

biological model.

 Simultaneous generation of static and dynamic magnetic fields.

 Opportunity of varying magnetic field direction and generating linearly

and circularly polarized fields.
ELF exposure systems for in vitro and in vivo experiments

• How to generate a uniform magnetic field?

ELF exposure systems for in vitro and in vivo experiments

Generation of static and ELF magnetic field

Static and ELF magnetic field can be generated by:

only
 Permanent magnets and ferromagnetic materials static
field
 Ferromagnetic structures with loop of wires

 Other special arrangements: Crawford cell*, flat plates**

 Current loops

*(Spiegel et al. 1987; Paul et al. 1995)

**(Gundersen 1986; Miller 1989)

 ELF exposure systems for in vitro and in vivo experiments

Generation of magnetic field at ELF: ferromagnetic

material with loops
“Air-gap reactor constructed from lamination of a
ferromagnetic material with a multiturn winding of
wire on one arm and an air gap in the opposite one.
A rectangular medium filled chamber containing living
cells is placed in the air gap.”

N=1000; lgap= 0.015 m

! B > 80 mT with 1 A
NI B = 8.316 10-2 T for one Ampere
B = µ0 of current applied to the coil
gap ! length
Reference:
Mullins RD, Sisken JE, Hejase HAN and Sisken BF: ’Design and Characterization of a System for Exposure of Cultured
Cells to Extremely Low Frequency Electric and Magnetic Fields Over a Wide Range of Fields Strengths’
Bioelectromagnetics 14: 173-186, 1993.
 ELF exposure systems for in vitro and in vivo experiments

Generation of magnetic field at ELF: MF produced by

“sheets of currents”
Three parallel plates used in vertical stacked configuration and appropriately energized:
the sheet of current in the central plate divides equally between the upper and the lower ones
producing a magnetic field, parallel to the plates, that is in opposite direction in the two zones

I/2

B E
I
V- V+
I/2
B
E I
I/2 I/2

Magnetic field lines

! These kind of systems result inadequate to the present research needs in the
bioelectromagnetic area that requests high homogeneous field volumes
Reference:
Miller DL, Miller MC and Kaune WT, “Addition of Magnetic Field Capability to Existing Extremely Low Frequency Electric Field
Exposure System”, Bioelectromagnetics 10: 85-98, 1998.
ELF exposure systems for in vitro and in vivo experiments

Generation of magnetic field at ELF: MF produced by

“sheets of currents”
An equivalent systems consist of two
I/2 rectangular solenoid…
I
I/2

Magnetic field lines

This arrangements eliminates

significant skin effects and
requires less current than I/2
B
‘plates’ to produce the same
I/2
field
B

! These kind of systems result inadequate to the present research needs in the
bioelectromagnetic area that requests high homogeneous field volumes
ELF exposure systems for in vitro and in vivo experiments

Generation of magnetic field at ELF: system of loops

square loops
z z-component of flux density B (due to one coil) at the
2b location of P is the sum of the contribution of each side (α)

2a
r’2 NI
r’1 Bzi = µ 0 ! f (r " " )
P(x,y,z)
4# "

s By the principle of superposition, the flux density at P due

r3 to the current in both loops (assuming the same N and
r2
r1 r4
current direction), is:

y
Bz (P) = !B zi (rP )
x
i

Because of symmetry consideration, Bx and By vanish along the z-

axis and the total flux density B is equal to the z-component Bz
ELF exposure systems for in vitro and in vivo experiments

Generation of magnetic field at ELF: system of loops

square loops: 1.2
Bz along vertical line
Graphs show Bz(x,y,z) / Bz(0,0,0.1) at points (x, y ≠ 0)
1.1
• the uniformity of Bz in a
wide area (with respect to Bz along z-axis

Normalized flux density

1.0
the overall volume),
• Bx and By components are Bz(0,0,z) / Bz(0,0,0.1)
0.9
negligible in the same
z/s
area. 0 0.25 0.5 0.75 1.0

0.2

Bx and By along vertical

0.1 line at points (x, y ≠ 0)
Bx(x,y,z) / Bz(0,0,0.1)
By(x,y,z) / Bz(0,0,0.1)
0

-0.1

z/s
0 0.25 0.5 0.75 1.0
ELF exposure systems for in vitro and in vivo experiments

Generation of magnetic field at ELF: system of loops

circular loops
z
I z-of flux density B at the location of P is given by the
b following formula.

d P(r,θ) As for square loops, for symmetry consideration, Bx

and By vanish along the z-axis and the total flux
y density B is equal to the z-component Bz
x
d
I

NIb 2
Bz (r , !) = µ 0
2(b 2 + d 2 )1.5
[
2 + A P
2 2 (cos ! )r 2
+ A P
4 4 (cos ! )r 4
+ ... ]
where: A2 = f ' (b, d ) [m-2] and A4 = f '' (b, d ) [m-4]
ELF exposure systems for in vitro and in vivo experiments

Generation of magnetic field at ELF: solenoids

[In a solenoid N turns of wire wound on a cylindrical form of length L and radius a]

L
NI
a
Bz ( z ) = µ 0 (cos !1 + cos !2 )
θ1 θ2 2L
z
P NI
Bz (L / 2) = µ 0
4a 2 + L2
z

For a long cylinder (L>>a) the field within the solenoid is nearly uniform
except close to the ends and to the turns of wire …
ELF exposure systems for in vitro and in vivo experiments

Generation of magnetic field at ELF: ferromagnetic

materials or coils?
Systems using ferromagnetic core
 are able of producing UNIFORM magnetic flux density across a wide range of
values (up to some Teslas), higher than those generated with systems of coils of
similar dimension
 cannot vary the magnetic vector direction or generate circularly polarized field

Systems realized by coils and loops

 are restricted in generating high magnetic field by the currents that flow
through the wires …

 can vary the magnetic vector direction and generate circularly polarized field
 can generate and control simultaneously both the static and dynamic field
components
ELF exposure systems for in vitro and in vivo experiments

Most suitable arrangements for the generation of

uniform magnetic fields and for satisfying other
fundamental requirements are

systems of coils
ELF exposure systems for in vitro and in vivo experiments

Steps and improvements in the generation of the uniform

magnetic fields with coils

I. Two axial coils (Helmholtz) Increasing the level of generated field

II. Set of more than two coils: Increasing the level of generated field
• Rubens (1945) and improving the high uniformity
volume
• Lee-Whiting (1957)
• Alldred & Scollar (1967)
• Merritt (1983)
III. Double wrapped coil Generation of null field

IV. Multiple orthogonal sets Generation of circularly polarized fields

and anyway oriented field
 ELF exposure systems for in vitro and in vivo experiments

Basic concepts and definitions I

set
LOOP: single circular or square wire
COIL: several turns of wire
SET: several axial coils (usually from 2 to 5)
B
SYSTEM: One, two or three orthogonal sets

loop
coil coil section

B
B Coils (circular)

wires

Scheme of two orthogonal sets of two

coils (multiwire) each, for the
generation of circularly polarized
magnetic field
 ELF exposure systems for in vitro and in vivo experiments

Basic concepts and definitions I

Generation of low intensity magnetic field at ELF: one single coil

B I
loop B = µ 0H = µ 0 (T)
2r

NI
coil
x
B = µ 0H = µ 0 (T)
B z 2r

N: number of turns of the coil;

I: current which flows through it (A)
r: coil radius (m)
 ELF exposure systems for in vitro and in vivo experiments

Basic concepts and definitions I

Generation of magnetic field at ELF: two coils (Helmholtz)

C1 C2 Using two axial coils the field in the center of the system can
be increased …
It is the sum of the contributions of the two coils
r

Px C
[
B = B z = µ 0Hz|( y =0 ) = µ 0 HCz1 + HCz2 ]
x

' $
NIr 2 ! 1 1 !
Hz(y = 0 ) = & + #
2 ! 2 ( [ )2
]
3
[r 2 2
]
3
a
% r + a ( x + (a + x ) !"
a = r/2

x=0
a = r/2

NI NI
Hz(y = 0 ) = 0,7155 = 1,43
r 2r
ELF exposure systems for in vitro and in vivo experiments

Basic concepts and definitions I - Helmholtz system

• Most common commercial generators have R=50 Ω and Vmax<40V …
One coil (radius r, N turns, …) Two coils (radius r, N turns, …)
y C1 C2
N is increased by a factor 2
r
x I is divided by a factor 2
B
x R required for each coil is 100Ω
B z
(the 2 coils are set in parallel)
a
a = r/2

NI NI
B = µ 0H = µ 0 B = 1,43µ 0 (T)
2r 2r

Using the same generators (R=50 Ω; Vmax<40V)

we can increase the intensity field and the
uniformity area
ELF exposure systems for in vitro and in vivo experiments

How design a system of coils?

Which factors should be considered?

ELF exposure systems for in vitro and in vivo experiments

• uniformity flux density volume (as a function of D)

• field exposure level (required B)
are defined by the experiment design
 ELF exposure systems for in vitro and in vivo experiments

Definitions of technical parameters

Designing ELF exposure systems several geometrical and physical

constrains should be considered.

These constrains concern, mainly, the following physical factors:

a) Small-diameter coils do not allow large spatial region of uniformity
b) Large-diameter coils cause weight and cost problems
c) High value of current cause high power dissipation and temperature
increase
 ELF exposure systems for in vitro and in vivo experiments

Definitions of technical parameters

Geometrical and physical constrains
wire
Elements definition: t/2 t/2
d - is the wire diameter
d t - is the coil thickness
D – is the coil diameter (2r)
Coil cross-section D/2

Geometrical constrain:
 Maximal distance between coils is D/2
 Minimal distance … is t=√N d
Nd < D / 2
t = Nd

Physical constrain:
2
l N"D ND 't$ R d2
R=! =! 2
= 4! 2 N=% " =
S d d &d # 4! D
"
4 If we want to keep R=100 Ω …
N is only a function of D and d
ELF exposure systems for in vitro and in vivo experiments

How determine optimal values for the

coil diameters (D),
wire diameter (d),
number of turns (N),
current (I) ?
 ELF exposure systems for in vitro and in vivo experiments

Definitions of technical parameters

Geometrical and physical constraints: number of turns N and B level

Example: Field intensity B (mT) as a function of coil diameter D and wire diameter d with 10 V
D (mm)
Required 0.2 0.25 0.3 0.4 0.5 0.6 0.85
1/ 2
D ≥ 15 cm 30 11.8 a 18.4 a 26.4 a 47.0 a 73.4 a 106 a 212 a &R# d2
and 50 4.23 6.61 a 9.52 a 16.9 a 26.4 a 38.1 a 76.4 a t = $$ !!
% 4' " D1 / 2
B = 1 mT 100 1.06 1.65 2.38 4.23 b 6.61 a 9.52 a 19.1 a
150 0.47 0.73 1.06 1.88 2.94 b 4.23 a 8.49 a
180 0.33 0.51 0.73 1.31 2.04 2.94 b 5.89 a
a: unreasonable value;
200 0.26 0.41 0.59 1.06 1.65 2.38 4.77 a b: acceptable value

Coil thickness t (mm) as a function of coil diameter D and wire diameter d for 100 Ω
D (mm)
0.2 0.25 0.3 0.4 0.5 0.6 0.85
30 8.9 13.8 19.9 a 35.4 a 55.3 a 79.7 a 159.9 a
50 6.9 10.7 15.4 27.4 b 42.9 a 61.7 a 123.8 a
100 4.8 7.6 10.9 19.4 30.3 43.6 87.6 a
' t2 $
150 4.6 6.2 8.9 15.8 24.7 35.6 71.5 N = %% 2 "" ! 880
180 3.6 5.6 8.1 14.5 22.6 32.5 65.3 &d #
200 3.4 5.4 7.7 13.7 21.4 30.9 61.9 ' t2 $
N = %% 2 "" ! 1180
&d #
Reference:
De Seze R., Lahitte A., Moreau J.M., Veyret B.:’ Generation of extremely-low frequency magnetic fields with standard
available commercial equipment: implications for experimental bioelectromagnetics work’. Bioelectrochemistry and
Bioenergetics 35: 127-131, 1994.
 ELF exposure systems for in vitro and in vivo experiments

Definitions of technical parameters

Feasibility: current density

Coil section, length and mass as a function of wire diameter (copper, R=100 Ω)
coil _ length = N!D Wire diameter
d (mm)
Section
S (mm2)
Length
l (m)
Mass
M (g)
0.2 0.03 185 52

coil _ length 0.3 0.07 416 261

=k 0.4 0.13 739 826
Length 0.5 0.20 1154 2016
0.6 0.28 1662 4181
0.85 0.57 3336 16841
System Mass = 2 · k · Mass
1 0.79 4618 32261a
2 3.14 18471 516179b

Reference:
De Seze R., Lahitte A., Moreau J.M., Veyret B.:’ Generation of extremely-low frequency magnetic fields with standard
available commercial equipment: implications for experimental bioelectromagnetics work’. Bioelectrochemistry and
Bioenergetics 35: 127-131, 1994.
 ELF exposure systems for in vitro and in vivo experiments

Definitions of technical parameters

Feasibility: current density
1 B
I = 2r
1.43µ 0 N
Current density J (A/mm2) as a function of wire diameter and section
I (A) 0.2 mm 0.3 mm 0.4 mm 0.5 mm 0.6 mm 0.85 mm 1 mm 1.55 mm
0.03 mm2 0.07 mm2 0.13 mm2 0.20 mm2 0.28 mm2 0.57 mm2 0.79 mm2 1.77 mm2

0.10 3.2 1.4 0.80 0.51 0.35 0.18 0.13 0.06

0.20 6.4 2.8 1.6 1.0 0.71 0.35 0.25 0.11
0.50 16 7.1 4.0 2.5 1.8 0.88 0.64 0.28
1.00 32 14 8.0 5.1 3.5 1.8 1.3 0.6
2.00 - 28 16 10.2 7.1 3.5 2.5 1.1
5.00 - - 40 25.5 17.7 8.8 6.4 2.8
10.00 - - - - 35.4 17.6 12.7 5.7
20.00 - - - - - 35.3 25.5 11.3
50.00 - - - - - - - 28.3
Acceptable values Unreasonable values

Reference:
De Seze R., Lahitte A., Moreau J.M., Veyret B.:’ Generation of extremely-low frequency magnetic fields with standard
available commercial equipment: implications for experimental bioelectromagnetics work’. Bioelectrochemistry and
Bioenergetics 35: 127-131, 1994.
ELF exposure systems for in vitro and in vivo experiments

Improvements in the generation of

uniform magnetic fields with coils
ELF exposure systems for in vitro and in vivo experiments

• Helmholtz configuration consists of a pair of parallel coils separated at a

distance obtained with the condition that the first and second spatial
derivatives of the applied field are equal to zero at the center of volume
between the two coils.

• Subsequent work showed that several higher-order derivatives can be

zeroed using assemblies of three, four or five coils, yielding much larger
volumes of uniform field space.

• From this concept several systems of different size, coils shape, numbers
of coils and wires have been studied and performed by several
researchers.
ELF exposure systems for in vitro and in vivo experiments

Basic “reference” systems

A. - Helmholtz

B. - Rubens

C. - Lee-Whiting

D. - Merritt et al.

E. - Merritt et al.

F. - Alldred & Scollar

 ELF exposure systems for in vitro and in vivo experiments

Design specifications for basic “reference” systems

Central
Coil N° of Coil diameter Coil spacing w.r.t. Ampere-turn
field
Shape coils or side length center of system Ratios
(µT/A)

circular 2 D, D -0.25 D, +0.25 D 1/1 1.798/D

Helmholtz
square 2 D, D -0.27 D, +0.27 D 1/1 1.629/D

-0.47 D, -0,12 D,
Lee-Whiting circular 4 D, D ,D ,D 9/4/4/9 17.96/D
+0.12 D, +0.47 D

Merritt et al. -0.41 D, 0,

(1) square 3 D, D, D 39 / 20 / 39 68.21/D
+0.41 D

Merritt et al. -0.51 D, -0.13 D,

(2) square 4 D, D ,D ,D 26 / 11 / 11 / 26 46.65/D
+0.13 D, +0.51 D

Alldred and 0.95 D, D, -0.52 D, -0.14 D,

square 4 21 / 11 / 11 / 21 40.29/D
Scollar D, 0.95 D +0.14 D, +0.52 D

-0.5 D, -0.25 D, 0,
Rubens square 5 D, D ,D ,D, D 19 / 4 / 10 / 4 / 19 35.69/D
+0.25 D, +0.5 D

Reference:
Kirschvink JL, ‘Uniform Magnetic Field and Double Wrapped Coil Systems: Improved techniques for the design of bioelectromagnetics
experiments’, Bioelectromagnetics 13: 401-411 (1992)
ELF exposure systems for in vitro and in vivo experiments

Uniformity levels of basic coil configurations

Helmoltz
High uniformity area:
20 cm x 20 cm
Area within 20% contour:
1m x 1m

Merritt et al. (4 coil) –

Alldred & Scollar
High uniformity area:
40 cm x 40 cm
Area within 20% contour:
1,4 m x 1,4 m
 ELF exposure systems for in vitro and in vivo experiments

Overall dimensions and uniformity of basic “reference” systems

Overall Uniformity region Uniformity region Uniformity region

Dimension 1% 0.1% 0.01%

Helmholtz 0.393 D3 0.049 D2 0.016 D2 0.006 D2

Lee-Whiting 0.739 D3 0.371 D2 0.212 D2 0.067 D2

Merritt et al. (1) 0.821 D3 0.169 D2 0.096 D2 0.046 D2

Merritt et al. (2) 1.01 D3 0.132 D2 0.163 D2 0. 086 D2

Alldred and
0.984 D3 0.228 D2 0.147 D2 0.04 D2
Scollar

Rubens D3 0.269 D2 0.115 D2 0.012 D2

Reference:
* Gottardi G, Mesirca P, Agostini C, Remondini D and Bersani F, ‘A Four Coil Exposure System (Tetracoil) Producing a Highly Uniform
Magnetic Field’, Bioelectromagnetics 24: 125-133 (2003)
ELF exposure systems for in vitro and in vivo experiments

Last steps for the realization of the “ideal” system:

• Generation of null field: “double-wrapped coil”
• Generation of circular polarization: orthogonal sets
ELF exposure systems for in vitro and in vivo experiments

Why a null field should be generated

“Control experiments should be designed such that the only difference between
treated and untreated groups is the magnetic field …”
In addition to the magnetic field, the coils introduce other undesired changes
which might have some influence:
 Electrical (ohmic) heating effects (temperature increase)
 Variable-frequency noise and / or vibration
 Small electric fields produced by the voltage drop between loops within
the coils

The proposed solution consist in the realization of coils “wrapped in parallel

with two separate strands of wire” … where currents can flow in parallel and
in antiparallel direction …
Double-wrapped coils also facilitate the use of truly double-blind procedure.
 ELF exposure systems for in vitro and in vivo experiments

Generation of null field: basic technique

N
B = µ0 2
(I1 + I2 ) Double-wrapped coils
2r
Phase shifter
B’=2B
(0°)
I1= I2 : in out out

NI
B = µ0
2r
Double wrapped coil

Phase shifter B’= + B - B = 0

(180°)
I1= -I2 : in out out
N
B = µ0 2
(I1 + I2 ) = 0
2r
Double wrapped coil
ELF exposure systems for in vitro and in vivo experiments

Generation of circularly polarized field

Set 1 Magnetic field coils

Phase shifter
90°
in out 0°

B1
Set 2
B2

amplifiers

 Each set of coils generates a field components along its axis;

 the magnetic field vector is the sum of these components: it rotates and traces an ellipse
in the plane perpendicular to both sets;
 the period of rotation coincides with the period of the ac voltage applied to the loops.
ELF exposure systems for in vitro and in vivo experiments

Generation of oriented field

Set 1 Magnetic field coils

in out out
B
B1
Set 2
B2

amplifiers

 Each set of coils generates a field components along its axis;

 the magnetic field vector is the sum of these components: resulting B vector have the
desired direction
ELF exposure systems for in vitro and in vivo experiments

Design specifications for other systems

n° of coil diameter or lenght Number of sets Ampere-turn Basic

Coil Shape
coils /set (m) (polarization) Ratios configuration

Baum et al. Hor: 2.14 68 / 34 / 68

Square 3 2 (circular) Merrit at al. (1)
(1991) Ver: 1.96 64 / 32 / 64
Shigemitsu et Hor: 1.7
Square 5 2 (circular) 11 / 2 / 5 / 2 /11 Rubens
al. (1993) Ver: 1.8
Yasui & Double
4 3.15 x 1.16 1 (linear) 4/2/2/4 Harvey (*)
Otaka (1993) rectangular
X: int 1.099, ext 1.15
Raganella et 3 (linear and Alldred and
Square 4 Y: int 0.908, ext 0.95 21 / 11 / 11 / 21
al. (1994) Z: int 0.745, ext 0.78
circular) Scollar

Gottardi et al.
Circular 4 0.67 D / D / D / 0.67 D 1 (linear) 73 / 107 / 107 / 73 -
(2003)
ELF exposure systems for in vitro and in vivo experiments

Some recent systems for in vivo and in vitro experiments

ELF exposure systems for in vitro and in vivo experiments

An In vivo triaxial exposure system*

This tri-axial exposure system

provides static and low-frequency
magnetic fields with different
polarization.

* Raganella L, Guelfi M and d’Inzeo G, ‘Triaxial Exposure System Providing Static And Low-
frequency Magnetic Fields For In Vivo And In Vitro Biological Studies’, Bioelectrochemistry and
Bioenergetics 35 (1994)
ELF exposure systems for in vitro and in vivo experiments

In vivo triaxial exposure system

• The apparatus consists of three sets of four square coils (Alldred &
Scollar) to provide greater uniformity.
A magnetic field linearly polarized along the coil axis is allowed. Therefore, we need
three nested systems, each fed independently, to control the component directed along
its axis.
Such a configuration also generates elliptically polarized fields.
• Each coil is made by wrapping together two copper wires (Φ=2.5 mm)
on a fibreglass frame;
• If the current flows in the same direction in both wires the induced magnetic fields
add together producing the desired exposure conditions, while equal and opposite
currents produce a null field in the direction of each system;
• The untreated group can be placed in the same experimental conditions as the
treated group except for the presence of the magnetic field.

• The number of turns is 44 for the inner coils and 84 for the outer coils.
The carrying structure of the system is made entirely of plastic material;
ELF exposure systems for in vitro and in vivo experiments

In vivo triaxial exposure system

Isofield regions for the xz plane of this system (side D<1.2 m)

Uniformity levels are, from the inner to the outer region,

0.01%, 0.1% and 1% with respect to the central value.
DEFINITION
Uniformity of 1% > (50 x 70) cm2
with respect to the
central value:
z (m)
Bxz " B00
! 0.01
B00

x (m)
ELF exposure systems for in vitro and in vivo experiments

In vivo triaxial exposure system

Supply and control system:

1 1= triaxial system;
2=PC and Labview (NI);
3= analog output board*;
4=data acquisition board;
5=amplifiers;
8 7 6
6=digital teslameter;
7=triaxial magnetic field sensor and Hall effect
5 5 5 4 9 probe;
8=thermometer;
9=GPIB-PCIIA interface board (NI)
3
2

*The waveforms, one for each axis component, are generated in digital form (2) by
a D/A converter (3); the amplifiers (5) connect the board to the exposure system (1).
ELF exposure systems for in vitro and in vivo experiments

In vivo triaxial exposure system

• Field values generated by each system in the

center of the exposure area as a function of 1
the current
Bx/I= 140.129 µT/A
By/I= 169.629 µT/A 8 7 6
Bz/I= 206.600 µT/A
5 5 5 4 9
The control program compares the measured and the
desired values, and adjusts the waveform in order to
obtain a stable static component of the field along each
axis. 3
The dynamic components can be superimposed on the
static component.
2
The values of static and dynamic fields and the
temperature are shown on the panel.
ELF exposure systems for in vitro and in vivo experiments

Measurements of:
• background values (magnetic field) and
• residual values when nullifying the field
are necessary for planning the experiments
 ELF exposure systems for in vitro and in vivo experiments

General requirements: background measurements I

Electric and magnetic fields measurements: In vivo ELF exposure systems
Background Magnetic field
Background
Null magnetic field
magnetic field By
B=0, ¦I¦ = 11 A (µT)
B=0, I=0 (µT) Bx
A - 0,30
E
B 0,02 0,25 A B C
D
C - 0,40
D - 0,40
Bz
E - 0,20

Background Electric field

B=0 B=0
B = 1.0 mT B = 1.5 mT
I=0 ¦I¦ = 11 A
E (V/m) 3,5 16 20 30

The electric field is measured by Holaday HI3638 (directional sensor), while flux magnetic density by Holaday 3627 (isotropic sensor).
The static geomagnetic field is about 40 ±1 µT and has been measured by Group3-Danfysik-Hall sensor (directional) and Bartington
fluxgate magnetometer-MAG-01H (directional).
ELF exposure systems for in vitro and in vivo experiments

In vitro triaxial exposure system

A similar exposure system has been realized for in vitro experiments, scaling the
previous one.
ELF exposure systems for in vitro and in vivo experiments

General requirements: background measurements II

Electric and magnetic fields measures: In vitro ELF exposure systems

Square coils system (side 40 cm) is placed in a water jacket incubator.

Background Magnetic field

Background Null magnetic
magnetic field field By
B=0, I=0 (µT) B=0, ¦I¦ = 5 A (µT) Bx
B min 1,0 1,4 E
A B C
B max 1,4 1,8 D

Background Electric field Bz

B=0 B=0
B = 1,0 mT
I=0 ¦I¦ = 5 A
E (V/m) 3,5 27 31
ELF exposure systems for in vitro and in vivo experiments

An In vitro (Tetracoil) exposure system**

A particular four coils system in which the

coils are geometrically constrained on a
sphere, producing a very uniform magnetic
field over a wide volume.

** Gottardi G, Mesirca P, Agostini C, Remondini D and Bersani F, ‘A Four Coil Exposure System
(Tetracoil) Producing a Highly Uniform Magnetic Field’, Bioelectromagnetics 24: 125-133 (2003)
ELF exposure systems for in vitro and in vivo experiments

In vitro “Tetracoil” exposure system

Uniformity level within 1% (a) and 0.1% (b) with respect to the central value.

1%

a1 a2 a2 a1

-b1 -b2 b2 b1

R 0.1%

4 1

3 2
ELF exposure systems for in vitro and in vivo experiments

Use of shielding materials (µ-metal)

If the background values are significantly high compared to the planned

exposure level (B required by experiment design) or
exposed and sham-exposed groups need to be placed in the same
incubator
the system should be surrounded by µ-shield
thus
it should also be provided by forced ventilation.
ELF exposure systems for in vitro and in vivo experiments

In vivo experiments with their relative exposure systems

Flux density ;
Animals End-points Polarization Exposure system
frequency

Repacholi et al. Em-PIM1 1, 100, 1000 mT, Linear Two systems of four rectangular Helmoltz
Lymphoma induction
(1998) transgenic mice sham; 50 HZ (horizontal) coils each one, assembled coaxially

Mevissen, Incidence and growth of

Sprague-Dawley 0.3-1 mT, 30 mT, Solenoidal coils
Loscher et al. female rats
mammary tumor after
sham; 50 Hz
Linear
(dia= 40 cm; length= 66 cm)
(1993) treatment with DMBA

Incidence and growth of

Baum, Mevissen Sprague-Dawley 100 mT, Four coils Merrit system
mammary tumor with and Linear
et al. (1994) female rats sham; 50 Hz (coil length= 1 m)
without DMBA treatment

Incidence and growth of

Three orthogonally systems of four square
Marino et al. C3HxDBA implanted mammary 2 mT, Linear and
coils each one; basic configuration:
(1995/98) female mice tumor; comparison with X- sham; 50 Hz circular
Alldred & Scollar
rays treatment

EF: 65 kV/m, EF: two horizontal electrodes;

Rogers et al Behavioral and Vertical EF
Baboons MF: up to 100 mT; 60 MF: two sets of uniformly spaced
(1995) neuroendocrine effects
Hz
Horizontal MF
conductors, around the EF system

Lymphoma/leukemia
Swiss Webster 1 mT, sham;
Shen et al (1997) mice
induction on DMBA
50 Hz
Linear Three pairs of Helmoltz coils
treated animals
Three orthogonally systems of four square
Frasca et al C57BL/6 female 2 mT, sham; Linear and
Immune system function coils each one; basic configuration:
(1997) mice 50 Hz circular
Alldred & Scollar
EF: 1, 5, 25, 100 EF: two horizontal electrodes;
Margonato et al. Sprague-Dawley Various morphologic and Vertical EF
kV/m, MF: five pairs of vertically arranged
(1993/98) male rats chemical parameters
MF: 5, 100 mT; 50 Hz
Horizontal MF
rectangular coils
ELF exposure systems for in vitro and in vivo experiments

In vitro experiments with relative exposure systems

Flux density ;
Cells line End-points Polarization Exposure system
frequency

Harland & Inhibition of antiproliferative A rotating four coils Merrit system with
MCF-7 human breast 0.2, 1.2 mT; Horizontal or
Liburdy cancer cells
action of Tamoxifen and
60 HZ Vertical
a mu-metal chamber is placed in a
(1997) Melatonin commercial incubator

Coulton & Two pairs of Helmholtz coils

T-Lymphocytes from
Ca++ intracellular 20.9 mT; 16 Hz Horizontal and perpendicular to each other and four
Barker thymus of BALB-c
concentration 65.3 mT; 50 Hz Vertical small solenoid coils housed in a wooden
(1993) mice
incubator

Jurkat cells A pair of Helmholtz coils mounted on

Lindstrom et al. Intracellular calcium 0.04-0.15 mT
(human leukemic Vertical the objective stage of an inverted
(1995) oscillations 5-100 Hz
T-cell line) microscope

Primary pinealocytes Two pairs of Helmholtz coils

Rosen at al. Norepinephrine-induced 0.05 mT;
of Sprague-Dawley Vertical perpendicular to each other placed in a
(1998) production of Melatonin 60 Hz
rats mu-metal box inside an incubator

Two pairs of Helmholtz coils

Livingston at al. Human lymphocytes Cythogenetic and cell kinetic 0.22 mT;
Circular perpendicular to each other housed in an
(1991) and CHO fibroblasts studies 60 Hz
incubator
ELF exposure systems for in vitro and in vivo experiments

In vivo experiments

 Exposure systems for in vivo experiments require large uniform magnetic

field volumes since is fundamental to allow the simultaneous treatment of as
many animals as possible, thus configurations with square coils are generally
employed,
 for animal studies, it is suitable to generate circularly polarized magnetic
fields thus exposure apparatus with two orthogonal oriented systems of coils
are generally employed.
ELF exposure systems for in vitro and in vivo experiments

In vitro experiments

 The simple Helmholtz design provide an adequate field uniformity just for in
vitro experiments, and it’s widely employed for these ones, because is easier to
make for experiments that require relatively small volumes,
 apparatus with orthogonal oriented systems of coils would be employed also
for in vitro experiments, in order to allow the control of all the static and ELF
components.
ELF exposure systems for in vitro and in vivo experiments

References
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Bioelectromagnetics, Suppl. 1.
Barnes, F.S. (1996). Interaction of DC and ELF electric fields with biological materials and systems. Handbook of Biological Effects of
Electromagnetic Fields. Second Edition. C. Polk, E. Postow, eds., Boca Raton: CRC Press, 103-147.
Bassen, H., Litovitz, T., Penafiel, M. & Meister, R. (1992). ELF in vitro exposure systems for inducing uniform electric and magnetic
fields in cell culture media. Bioelectromagnetics, 13, 183-198.
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electric and magnetic fields. Bioelectromagnetics, 12, 85-99.
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