The Effect of Fluorescent Labelling on Deformability and Geometric Properties of Red Blood Cells...
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Transcript of The Effect of Fluorescent Labelling on Deformability and Geometric Properties of Red Blood Cells...
![Page 1: The Effect of Fluorescent Labelling on Deformability and Geometric Properties of Red Blood Cells Joanne Wong Supervisor: Dr. D. Jackson March 23,2011 Medical.](https://reader034.fdocuments.us/reader034/viewer/2022051517/5697bfe91a28abf838cb6eee/html5/thumbnails/1.jpg)
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
![Page 2: The Effect of Fluorescent Labelling on Deformability and Geometric Properties of Red Blood Cells Joanne Wong Supervisor: Dr. D. Jackson March 23,2011 Medical.](https://reader034.fdocuments.us/reader034/viewer/2022051517/5697bfe91a28abf838cb6eee/html5/thumbnails/2.jpg)
Acknowledgement
• Supervisor: Dr. Dwayne N. Jackson• Graduate Student Mentor: Baraa Al-Khazraji• Source of funding: NSERC
![Page 3: The Effect of Fluorescent Labelling on Deformability and Geometric Properties of Red Blood Cells Joanne Wong Supervisor: Dr. D. Jackson March 23,2011 Medical.](https://reader034.fdocuments.us/reader034/viewer/2022051517/5697bfe91a28abf838cb6eee/html5/thumbnails/3.jpg)
Introduction• Blood cells• Hemodynamic• Directly measure• Microspheres and fluorescent cells• Trace cells in microvasculature • Do stained cells behave the same as unstained
cells?
![Page 4: The Effect of Fluorescent Labelling on Deformability and Geometric Properties of Red Blood Cells Joanne Wong Supervisor: Dr. D. Jackson March 23,2011 Medical.](https://reader034.fdocuments.us/reader034/viewer/2022051517/5697bfe91a28abf838cb6eee/html5/thumbnails/4.jpg)
Hypothesis
There will be no differences in deformability or geometry between stained and unstained red blood cells
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Methods
Dye and incubate5 hours (FITC) Wash cells
1 hour
Extract blood
Hang test2 minutes
Image slides
Split portions
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Methods• Hang test
• ImageJ
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Theoretical Model
• Criteria for measurements
•
Circle diameter
Diameter
Thickness
(2)
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Theory
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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
![Page 10: The Effect of Fluorescent Labelling on Deformability and Geometric Properties of Red Blood Cells Joanne Wong Supervisor: Dr. D. Jackson March 23,2011 Medical.](https://reader034.fdocuments.us/reader034/viewer/2022051517/5697bfe91a28abf838cb6eee/html5/thumbnails/10.jpg)
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
![Page 11: The Effect of Fluorescent Labelling on Deformability and Geometric Properties of Red Blood Cells Joanne Wong Supervisor: Dr. D. Jackson March 23,2011 Medical.](https://reader034.fdocuments.us/reader034/viewer/2022051517/5697bfe91a28abf838cb6eee/html5/thumbnails/11.jpg)
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
![Page 12: The Effect of Fluorescent Labelling on Deformability and Geometric Properties of Red Blood Cells Joanne Wong Supervisor: Dr. D. Jackson March 23,2011 Medical.](https://reader034.fdocuments.us/reader034/viewer/2022051517/5697bfe91a28abf838cb6eee/html5/thumbnails/12.jpg)
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
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Complementary Study Methods
StainedUnstained
Albumin free Albumin0.5% per 500mL
Albumin 0.5% per 500mL
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Complementary StudyUnstained Stained
No Albumin ~5% abnormal shaped
Albumin ~95% abnormal shaped
~30% abnormal shaped
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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?