Some Effects of Electric and Magnetic Fields on the

Movement of White Blood CellsKalani 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

• 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 25-35%; Monocytes account for 3-9%.

• 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.

Eosinophil

Neutrophil Monocyte

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

Cell Separation Drawing Sample

Buffy Layer

Making the Slides

Small Needle

C-AMP SLIDE

Micro Pipette

Sample Drop Slide

Needle

Push Cover slipCover Slip

• Make C-AMP sample to known concentration (120 mM/L)

• Using a small needle, draw a tiny stripe of the C-AMP solution on the slide

• Place a small drop of the WBC sample on the slide

• Sample at least ½ cm away from the stripe• Cover Slip placed on sample without covering

the stripe• Use a needle to push the slip over the stripe until

it is fully covered.• Apply Vaseline around slip to keep moist

 

Typical concentrations as a function of time and distance from the strip.

White cell movement

Positive chemotaxis

1

2

3

4

5

6

78

9

10 11 12

13

14

White cell movementwithout the effect of RF

radiation,

Velocity

( µm/min)

1

10

C0 molar/l

10-4 5×10-3 10-2 5×10-2 10-1 5×10-1 1 5×10-5

Note: when chemoattractant concentration is located between 8×10-2 molar/l and 5×10-1 molar/l, the WBC velocity can reach maximum.

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Chemotatic Velocities

What a Data Sample Looks Like• Variables

– 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

Experiment 8 (overnight)      

C-AMP = 0.002g         Serum = 0.05ml      

Elec field = 12V/mm   8 Temp = 37C   Person : kalani 1    

D1 = D2= 1mm    

Points Up Down DistanceTime

(Mins)

1--2 2 1 5.590169944 74

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     0.764442919  

Velocity W/O field     3  

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 : SpeedField Strength

(V/mm)Speed With (µm/min)

Speed Without (µm/min)

DC ELECTRICFIELD

6.67 1.12 3.0

12 1.2 3.0

20 2 3.2

REVERSEDIRECTION

-12 2.65 3.0-20 3.3 3.32

AC ELECTRICFIELD (V/mm)

13.3 0.9 3.216.67 1.1 3.2

20 3.97 3.32

Data Analysis : Speed – AC Fields

• 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

Speed V AC Electric Field

0

1

2

3

4

5

12 14 16 18 20 22

AC Electric Field Strength (V/mm)

Spe

ed (M

icro

ns/M

in)

Speed w/ Field

Speed with Field v Speed without Field

0

1

2

3

4

5

12 14 16 18 20 22

AC Electric Field Strength (V/mm)

Spe

ed (M

icro

ns/M

in)

Speed w/ Field

Speed w/o Field

Data Analysis : Speed – AC v DC

• 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

Speed with AC Field v Speed with DC Field

0

1

2

3

4

5

12 14 16 18 20 22

Electric Field Strength (V/mm)

Spee

d (Mi

cron

s/Min)

Speed w/ AC fieldSpeed w/ DC field

Data Analysis : Direction of Motion

Without any exposure –75% move toward C-AMP–20% move against C-AMP–5% random motion

0

10

20

30

40

50

60

70

80

Percentage of Experiments

Positive Chemotaxis NegativeChemotaxis

Random Motion

Direction of Motion without Exposure to any Fields

Changes in the direction of motion: DC Fields

• Often when exposed, cell changes its direction of motion

• Not necessary towards the field but to a different direction from its initial

• 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

1

1019

36

C-AMP

E

1 – 10 Without E Field

10 – 19 With E Field

19 – 36 Reversed E Field

Statistics for DC Fields

• 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

01020304050607080

Percentage

Changeddirection of

motion w/ field

Changeddirection withreversed field

Reverseddirection w/

reversed field

Became moreactive when field

removed

Change in Direction due to Exposure

Statistics for AC Fields

• 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

010203040506070

Percentage

Move in anarc w/ field

Stoppedafter a

while ofexposure

Movedrandomlywhen fieldremoved

Becamemore activewhen fieldremoved

Change in Direction due to Exposure : AC Fields

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°

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

1

9

14

19

BDC( µT) Velocity (µm/min)

w/o BDC with BDC w/o BDC again

15 4.11 4.93 4.69

22.89 5.02 6.83 5.81

25.33 3.22 3.68 2.79

31.2 3.89 6.8 5.42

35.73 4.65 5.13 4.87

39.2 3.0 4.40 3.37

52.27 5.78 6.65 2.06

63.47 4.58 6.48 4.50

71.43 3.24 4.38 2.85

78.4 5.65 7.34 6.15

101.43 3.03 4.34 2.13

DC magnetic field versus WBC velocity

Low frequency AC magnetic field effect on WBC mobility

Experiment 1: Co=1.2E-1 molar/l; B=18.67µT; T=38°; f=60Hz

Results: 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=2.12m/min

Note: The WBC move toward chemoattractant

Chemoattractant gradient

1

8

15

19

BAC( µT) Velocity (µm/min)

w/o BAC with BAC w/o BAC again

11.2 4.21 4.29 (60Hz) 5.87

6.7 5.28 (25Hz) 5.94

6.23 5.77 (15Hz)

14.67 4.04 3.51 (60Hz) 3.31

5.29 3.65 (60Hz)

18.67 5.84 4.13 (60Hz) 2.16

3.88 2.88 (60Hz) 2.12

20.7 6.90 5.04 (5Hz) 4.36

2.37 2.15 (5Hz) 3.3

AC magnetic field versus WBC velocity

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)

Positive chemotaxis

2

3

4

5

6

7

8

9

1

RF

Cell movementunder RF(990MHz), which

shows the effect of RFradiation on the cellmovement, and also

shows how the effect ofRF radiation eliminate the

effect of C-Amp.

1-4 without RF4-9 under RF

Results

Without RF radiation

With RF radiation

Cell response time constant to RF radiation

The cells act normally

2.5 minutes

Av. Movement speed 2.4 μm/min 4.5 μm/min

Av. Chemotactic index 0.3 0.8

Changing shape Faster Movement direction Sideward direction

Upward direction

Preliminary Conclusion1. 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?