Comparison of immunophenotyping by slide-based cytometry and by flow cytometry

Journal of Immunological Methods
Andreas O H GerstnerAttila Tárnok

Abstract

Flow cytometry (FCM) is the gold standard for immunophenotyping of peripheral blood leukocytes (PBLs). Slide-based cytometry (SBC) systems, for example the laser scanning cytometer (LSC(R), CompuCyte), can give additional information (repeated staining and scanning, morphology). In order to adequately judge the clinical usefulness of LSC for immunophenotyping it is obligatory to compare it with FCM. The aim of this study was to systematically compare immunophenotyping by both FCM and LSC methods and to test the correlation of the results. PBLs were stained with directly labeled monoclonal antibodies with the whole blood staining method. Aliquots of the same paraformaldehyde fixed specimens were analyzed in parallel by a FACScan (BD-Biosciences) using standard protocols and by LSC with different triggers (forward scatter, CD45 FITC, or 7-AAD). For 7-AAD measurements by LSC, slides were additionally fixed with acetone before 7-AAD staining. Calculating the percentage distribution of PBLs obtained by LSC and by FCM showed very good correlation with regression coefficients close to 1.0 for the major populations and the lymphocyte sub-populations (neutrophils, monocytes, and lymphocytes; T-helper-, T-cytotoxic-, B-, NK-cells). The b...Continue Reading

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Citations

Mar 21, 2008·HNO·A O H Gerstner, W Laffers
Sep 13, 2011·Apoptosis : an International Journal on Programmed Cell Death·Leo Li-Ying ChanBo Lin
Jun 22, 2012·Journal of Fluorescence·Leo L ChanNing Lai
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