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