the effect of fluorescent labelling on deformability and geometric properties of red blood cells...

The Effect of Fluorescent Labelling on Deformability and Geometric

Properties of Red Blood Cells

Joanne WongSupervisor: Dr. D. Jackson

March 23,2011Medical Biophysics Department University of

Western Ontario

Acknowledgement

• Supervisor: Dr. Dwayne N. Jackson• Graduate Student Mentor: Baraa Al-Khazraji• Source of funding: NSERC

Introduction• Blood cells• Hemodynamic• Directly measure• Microspheres and fluorescent cells• Trace cells in microvasculature • Do stained cells behave the same as unstained

cells?

Hypothesis

There will be no differences in deformability or geometry between stained and unstained red blood cells

Methods

Dye and incubate5 hours (FITC) Wash cells

1 hour

Extract blood

Hang test2 minutes

Image slides

Split portions

Methods• Hang test

• ImageJ

Theoretical Model

• Criteria for measurements

•

Circle diameter

Diameter

Thickness

(2)

Theory

Results

n = 99n = 99

Mean value :Unstained= 7.624 ± 0.475 µmStained= 7.440 ± 0.367 µmA T-test was performed with p value = 0.179

Unstained Stained0

1

2

3

4

5

6

7

8

Diameter Value for Stained and Unstained Rat Blood Cells

Dia

mte

r (µm

)

n=20n=20

n = 99n = 99

Mean value :Unstained= 130.890 ± 15.711 µm2

Stained = 125.490 ± 12.739 µm2

A T-test was performed for a p value = 0.24

Unstained Stained0

20

40

60

80

100

120

140

Surface Area for Stained and Unstained Rat Blood Cells

Surf

ace

Are

a (µ

m2)

n=20n=20

n = 99n = 99

Mean value :Unstained= 48.544 ± 10.025 µm3

Stained = 46.158 ± 4.615 µm3

A T-test was performed with a p value = 0.340

Unstained Stained0

10

20

30

40

50

60

Volume of Stained and Unstained Rat Blood Cells

Volu

me

(µm

3)

n=20n=20

n = 99

Mean value :Unstained= 0.490± 0.022Stained = 0.498 ± 0.037A T-test was performed with a p value = 0.357

Unstained Stained0

0.1

0.2

0.3

0.4

0.5

Sphericity Index Value for Stained and Unstained Rat Blood Cells

Sphe

rici

ty In

dex

n=20n=20

Complementary Study Methods

StainedUnstained

Albumin free Albumin0.5% per 500mL

Albumin 0.5% per 500mL

Complementary StudyUnstained Stained

No Albumin ~5% abnormal shaped

Albumin ~95% abnormal shaped

~30% abnormal shaped

Summary & Discussion

• Simple math model vs. hand tracing• Results showed no significant difference• Effects of albumin buffer• FIT-C dye does not affect deformability and

geometrical difference• Possible modification of staining protocol?

Thank You!

Contact me at: twong227@uwo.ca