The Effect of Fluorescent Labelling on Deformability and Geometric Properties of Red Blood Cells Joanne Wong Supervisor: Dr. D. Jackson March 23,2011 Medical 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 incubate 5 hours (FITC) Wash cells 1 hour Extract blood Hang test 2 minutes Image slides Split portions

Methods Hang test ImageJ

Theoretical Model Criteria for measurements Circle diameter Diameter Thickness (2)

Theory

Results n = 99 Mean value : Unstained= ± µm Stained= ± µm A T-test was performed with p value = n=20

n = 99 Mean value : Unstained= ± µm 2 Stained = ± µm 2 A T-test was performed for a p value = 0.24 n=20

n = 99 Mean value : Unstained= ± µm 3 Stained = ± µm 3 A T-test was performed with a p value = n=20

n = 99 Mean value : Unstained= 0.490± Stained = ± A T-test was performed with a p value = n=20

Complementary Study Methods Stained Unstained Albumin free Albumin 0.5% per 500mL Albumin 0.5% per 500mL

Complementary Study UnstainedStained 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?

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