Lymphocyte subset determination using a hematology analyzer.
Anti-CD4 antibody (T4)-coated microspheres were used to label CD4 cells in whole blood. The mixture was lysed and analyzed by a modified Coulter VCS hematology analyzer, which differentiated microsphere-labeled cells by a change in Coulter volume, conductance, and light scatter. %CD3+/CD4+ fluorescent values from a profile were compared to %CD4 values using the VCS-microsphere method. CD3 gating was used to exclude CD4+ monocytes from the 90LS-FALS lymphocyte gate. The results correlated well (R = 0.996). The percentage of CD4+ lymphocytes from profile scatterplots and VCS scatterplots showed a line of regression close to the equivalence line (n = 76, slope = 0.96) when CD3 gating was used for the profile. These results suggest that CD3 gating, though necessary for 90LS-FALS scatterplots, may not be necessary for volume-conductance-light scatterplots.
Hudson
J C
JC
Coulter Corporation, Miami, FL 33116-9015, USA.
Brunhouse
R F
RF
Garrison
C
C
Rodriguez
C M
CM
Zwerner
R
R
Russell
T R
TR
D000918:Antibody Specificity; D001322:Autoanalysis / Q000295:instrumentation*; D001667:Binding, Competitive; D001782:Blood Donors; D048429:Cell Size; D004553:Electric Conductivity; D005260:Female; D005434:Flow Cytometry; D006679:HIV Seropositivity / Q000276:immunology*; D006405:Hematology / Q000295:instrumentation*; D006801:Humans; D008027:Light; D016131:Lymphocyte Subsets; D008297:Male; D008863:Microspheres; D012542:Scattering, Radiation