Some Effects of Electric and
Magnetic Fields on the
Movement of White Blood Cells
Kalani Rathnabharathi
Rong Zhou, Ashraf Aly
Muhammad Imran Cheema
Deepak Anchala, Ryan.Laterza
Frank Barnes
University of Colorado
Outline of Talk
1.Background
2.Methods
3. Effects of Low Frequency Electric Fields
4. Effects of Low Frequency Magnetic
Fields
5. Effects of RF Fields
6. Some Conclusions
Background
1. Concerns about Cancer and Power Lines
2. Concerns about Cell Phones
3. Possibility that the Immune System is
activated by low fields behaving as a stress
4. Earlier Work with Lasers
5. Need for a low cost project
Why we focus on white blood cells
Neutrophil
• White blood cells (WBC) have five
different types. Normally, neutrophils
account for 50-70%; Eosinophils account
for less than 5%; Basophils represent less
than 1%; Lymphocytes accounting for 2535%; Monocytes account for 3-9%.
Eosinophil
Monocyte
• WBC play important rolls in the body’s
immune system. All of them participate in
defense of the body against infections and
other foreign materials.
• It is an easy way to track WBC moving
trail which is affected by some
chemoattractants and their gradient.
Basophil
lymphocyte
White Blood Cells ( WBC) chemotaxis
Definition
Chemotaxis is the process by which white blood cells are
attracted and move towards a chemo-attractant. Neutrophils are
our body's first line of defense against bacterial infections.
After leaving nearby blood vessels, these cells recognize
chemicals produced by bacteria in a cut or scratch and migrate
toward the chemoattractant with considerable speed.
Methods
Blood collection
Centrifuge
Buffy Layer
Drawing Sample
Cell Separation
Making the Slides
• Make C-AMP sample to known concentration
(120 mM/L)
Small Needle
C-AMP
SLIDE
• Using a small needle, draw a tiny stripe of the CAMP solution on the slide
• Place a small drop of the WBC sample on the
slide
Micro Pipette
• Sample at least ½ cm away from the stripe
Sample Drop
Slide
• Cover Slip placed on sample without covering
the stripe
• Use a needle to push the slip over the stripe until
it is fully covered.
Needle
• Apply Vaseline around slip to keep moist
Cover Slip
Push Cover slip
Typical concentrations as a function of time and
distance from the strip.
White cell movement
Positive chemotaxis
3
White cell movement
without the effect of RF
radiation,
4
2
5
1
6
7
8
10
11
12
9
13
14
Chemotatic Velocities
10
Velocity
•
( µm/min)
•
1
•
•
•
5×10-5 10-4
•
•
5×10-3
•
•
10-2
•
• •
5×10-2
••
•
••
•••
•
10-1
5×10-1
1
C0 molar/l
Note: when chemoattractant concentration is located between 8×10-2
molar/l and 5×10-1 molar/l, the WBC velocity can reach maximum.
What a Data Sample Looks Like
• Variables
Experiment 8 (overnight)
C-AMP = 0.002g
Serum = 0.05ml
Elec field = 12V/mm
Temp = 37C
Person : kalani
8
1
D1 = D2= 1mm
Points
Up
Down
Time
(Mins)
Distance
1--2
2
1
5.590169944
2--3
4
2
11.18033989
3--4
4
2
11.18033989
4--5
3
2
9.013878189
5--6
2
0
5
6--7
3
2
9.013878189
7--8
1
2
5.590169944
56.56877604
Velocity
Velocity W/O field
0.764442919
3
74
– How old blood is
– Concentration of C-amp
– Electric field strength
– Temperature
– Person Blood sample belongs to
• Cell Movement
– Trace of cell movement
– Distance moved
– Time taken
– Speeds with and without field
Analysis Of Data
• Data analyzed in two categories: Speed & Direction
• Influence of fields on the speed of the cells
• Whether speed increases/ decreases with exposure compared with no
exposure
• Whether the speed increases/ decreases with the increase of field strength
• Whether there is a change in speed when field is reversed/removed
• Influence of fields on the direction of motion of the cells
• Do cells change their direction of motion once exposed to the fields
• Do cells reverse their direction when the fields are reversed
• What happens when the fields are removed
Few Data Samples : Speed
DC
ELECTRIC
FIELD
REVERSE
DIRECTION
AC
ELECTRIC
FIELD (V/mm)
Field Strength
(V/mm)
Speed With
(µm/min)
Speed Without
(µm/min)
6.67
12
20
-12
-20
13.3
16.67
1.12
1.2
2
2.65
3.3
0.9
1.1
3.0
3.0
3.2
3.0
3.32
3.2
3.2
20
3.97
3.32
Data Analysis : Speed – AC
Fields
Speed V AC Electric Field
Speed (Microns/Min)
5
4
3
2
1
0
12
14
16
18
20
22
AC Electric Field Strength (V/mm)
Speed w/ Field
Speed with Field v Speed without Field
Speed (Microns/Min)
5
4
3
Speed w/ Field
2
Speed w/o Field
1
0
12
14
16
18
20
AC Electric Field Strength (V/mm)
22
• Speed increase with field
• Exponential rise of speed
with respect to field
• Exponential rise is very
sharp
• Speed much lower than
without the fields for lower
field strengths
Data Analysis : Speed – AC v
DC
Speed with AC Field v Speed with DC Field
Speed (Microns/Min)
5
4
3
2
1
0
12
14
16
18
Electric Field Strength (V/mm)
20
22
Speed w/ AC field
Speed w/ DC field
• Less speed variations in DC
Fields
• Exponential rise in speed with
AC fields much higher than
with DC fields
• At lower field strengths both
DC and AC speeds are similar
Data Analysis : Direction of
Motion
Direction of Motion without Exposure to any Fields
Without any exposure
80
– 75% move toward C-AMP
– 20% move against C-AMP
– 5% random motion
70
60
Percentage of 50
Experiments
40
30
20
10
0
Positive Chemotaxis
Negative
Chemotaxis
Random Motion
Changes in the direction of
motion: DC Fields
• Often when exposed, cell changes
its direction of motion
C-AMP
E
36
1
19
1 – 10 Without E Field
10 – 19 With E Field
19 – 36 Reversed E Field
• Not necessary towards the field but
to a different direction from its
initial
10
• When field reversed, cell often
changes its direction again
• When field removed, cell often
continues in the same direction
• Sometimes when field is removed
cell becomes more active than
during exposure
Statistics for DC Fields
Change in Direction due to Exposure
Percentage
80
70
60
50
40
30
20
10
0
Changed
Changed
Reversed Became more
direction of direction with direction w/ active when field
motion w/ field reversed field reversed field
removed
•
With initial exposure
– 78% changed direction
• Reverse field
– 55% changed direction again
– 15% in reverse direction
• Remove field
– 30% became more active than
during exposure
Statistics for AC Fields
Change in Direction due to Exposure : AC Fields
Percentage
70
60
50
40
30
20
10
0
Move in an Stopped
Moved
Became
arc w/ field after a randomly more active
while of when field when field
exposure removed removed
• With initial exposure
– 67% cells moved in a
semi-circular shape
– 30% cells stop after a
while
• Remove field
– 58% moves randomly
– 45% became more active
than during exposure
Observation
• For some experiments it was observed that once the
direction of the DC field was reversed the cells also
switched its direction of movement
• However this results was not always reproducible
• Results vary from person to person and from day to
day
• Even if same person results vary with
– Level of exercise
– Foods taken
– Illnesses
DC magnetic field effect on WBC mobility
Experiment 1: Co=1.2E-1 molar/l; B=63.47µT; T=38°
1
9
14
Chemoattractant gradient
Results:
1-9 without DC magnetic field, v=4.58 m/min
9-14 with DC magnetic field, v=6.48 m/min
14-19 without DC magnetic field again, v=4.5 m/min
Note:
WBC move toward chemoattractant
19
DC magnetic field versus WBC velocity
BDC( µT)
Velocity (µm/min)
with BDC
4.93
15
w/o BDC
4.11
w/o BDC again
4.69
22.89
5.02
6.83
5.81
25.33
31.2
35.73
3.22
3.89
4.65
3.68
6.8
5.13
2.79
5.42
4.87
39.2
52.27
63.47
3.0
5.78
4.58
4.40
6.65
6.48
3.37
2.06
4.50
71.43
78.4
101.43
3.24
5.65
3.03
4.38
7.34
4.34
2.85
6.15
2.13
Low frequency AC magnetic field effect on WBC mobility
Experiment 1: Co=1.2E-1 molar/l; B=18.67µT; T=38°; f=60Hz
1
8
15
19
Results:
Chemoattractant gradient
1-8 without AC magnetic field, v=3.88 m/min
8-15 with AC magnetic field, v=2.88 m/min
15-19 without AC magnetic field again, v=2.12m/min
Note:
The WBC move toward chemoattractant
AC magnetic field versus WBC velocity
BAC( µT)
w/o BAC
Velocity (µm/min)
with BAC
w/o BAC again
11.2
4.21
6.7
6.23
4.29 (60Hz)
5.28 (25Hz)
5.77 (15Hz)
5.87
5.94
14.67
4.04
5.29
5.84
3.51 (60Hz)
3.65 (60Hz)
4.13 (60Hz)
3.31
3.88
2.88 (60Hz)
2.12
6.90
2.37
5.04 (5Hz)
2.15 (5Hz)
4.36
3.3
18.67
20.7
2.16
Conclusion
DC Magnetic field will
•Increase WBC velocity
•Make WBC move more randomly than without DC MF
•Change WBC direction of motion above 70µT
AC magnetic field will
•Decrease WBC velocity
•Change WBC direction of motion below 25Hz
White cell movement under
RF(990MHz)
9
Positive chemotaxis
Cell movement
under RF(990MHz), which
shows the effect of RF
radiation on the cell
movement, and also
shows how the effect of
RF radiation eliminate the
effect of C-Amp.
8
7
6
RF
3
5
1-4 without RF
4-9 under RF
4
2
1
Without RF
radiation
Cell response
time constant to
RF radiation
Av. Movement
speed
Av. Chemotactic
index
Changing shape
Movement
direction
With RF
radiation
The cells act
normally
2.5 minutes
2.4 μm/min
4.5 μm/min
0.3
0.8
Sideward direction
Faster
Upward
direction
Preliminary Conclusion
1. Leukocytes speed increased rapidly by raising temperature between 35°40°C (decreased above 40°C ).
2. Under the RF radiation, the movement speed will rise by about 50%.
3. Significant change in cells movement after exposure to RF radiation.
a. Cells movement direction , will be to the upward direction which is
perpendicular to the C-Amp direction direction (with no radiation effect
the cells moves sideward to the C-Amp direction ). No random
movements (which usually happened without the RF radiation effect).
b. Significant change in leukocytes behavior, include changing shape much
faster (about double the changing speed) than the normal case (without
applying RF radiation). The cells were shrinking, expanding, and rolling.
c. We got the same results by using mobile phone or signal generator
radiation.
d. The cells moves to the upward direction under the effect of RF radiation
in all tested temperatures Between 35° to 42°, and the speed of the cells
still depends on the temperature.
Some Topics to Explore
• 1. Thresholds for Effects
• 2. What are the induced Current Flows?
• 3. How does the cell process the signaling
information? Two Point? Integration over
time?
• 4. Is there a communication system between
leukocytes? IR?