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2014 ; 51
(2-3
): 159-70
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Deformability analysis of sickle blood using ektacytometry
#MMPMID24898336
Rabai M
; Detterich JA
; Wenby RB
; Hernandez TM
; Toth K
; Meiselman HJ
; Wood JC
Biorheology
2014[]; 51
(2-3
): 159-70
PMID24898336
show ga
Sickle cell disease (SCD) is characterized by decreased erythrocyte
deformability, microvessel occlusion and severe painful infarctions of different
organs. Ektacytometry of SCD red blood cells (RBC) is made difficult by the
presence of rigid, poorly-deformable irreversibly sickled cells (ISC) that do not
align with the fluid shear field and distort the elliptical diffraction pattern
seen with normal RBC. In operation, the computer software fits an outline to the
diffraction pattern, then reports an elongation index (EI) at each shear stress
based on the length and width of the fitted ellipse:
EI=(length-width)/(length+width). Using a commercial ektacytometer (LORCA,
Mechatronics Instruments, The Netherlands) we have approached the problem of
ellipse fitting in two ways: (1) altering the height of the diffraction image on
a computer monitor using an aperture within the camera lens; (2) altering the
light intensity level (gray level) used by the software to fit the image to an
elliptical shape. Neither of these methods affected deformability results
(elongation index-shear stress relations) for normal RBC but did markedly affect
results for SCD erythrocytes: (1) decreasing image height by 15% and 30%
increased EI at moderate to high stresses; (2) progressively increasing the light
level increased EI over a wide range of stresses. Fitting data obtained at
different image heights using the Lineweaver-Burke routine yielded percentage ISC
results in good agreement with microscopic cell counting. We suggest that these
two relatively simple approaches allow minimizing artifacts due to the presence
of rigid discs or ISC and also suggest the need for additional studies to
evaluate the physiological relevance of deformability data obtained via these
methods.