Comparative study of five commercial reagents for preparing normal and leukaemic lymphocytes for immunophenotypic analysis by flow cytometry
作者： M.G. Macey , D.A. McCarthy , T. Milne , J.D. Cavenagh , A.C. Newland
DOI: 10.1002/(SICI)1097-0320(19990815)38:4<153::AID-CYTO2>3.0.CO;2-E
关键词:
摘要: The flow cytometric analysis of leucocytes in whole blood is usually performed on samples which the erythrocytes have been lysed and fixed. Because lysis fixation reagents potential to introduce artefacts, several commercially available were used prepare normal leukaemic lymphocytes for immunophenotypic by cytometry, results compared with those obtained from live blood. tested ImmunoPrep system OptiLyse C (Coulter), LF-1000-Lyse Flow (Harlan), Uti-Lyse (Dako) FACS Lysing Solution (Becton Dickinson). effect each reagent apparent expression CD3, CD5, CD11b, CD45, FMC7, kappa lambda antigens was determined six controls patients chronic lymphocytic leukaemia (CLL). following observations made: (i) time minutes procedure varied markedly 1.5, 15, 20, 30 system, C, Uti-Lyse, Solution, LF-1000, respectively, but only 0.5 min (ii) forward side scatter characteristics affected all procedures, this most marked Flow. (iii) gave preparations poor resolution due presence residual red cell fragments. (iv) Lysis procedures did not affect or FMC7 CLL samples, highly variable kappa, samples. We conclude that can different artefacts are best avoided analysing
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