The Q-Prep system: Effects on the apparent expression of leucocyte cell surface antigens

✍ Scribed by M. G. Macey; D. A. McCarthy; C. Davies; A. C. Newland

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 77 KB
• Volume: 30
• Category: Article
• ISSN: 0196-4763
• DOI: 10.1002/(sici)1097-0320(19970415)30:2<67::aid-cyto1>3.0.co;2-a

✦ Synopsis

To facilitate the analysis of immunolabelled peripheral blood or bone marrow leucocytes by flow cytometry, a number of reagents are available commercially that lyse erythrocytes and fix leucocytes. This study has investigated the effect on antibody-labelled whole blood of the Q-Prep procedure, in which erythrocytes are lysed with formic acid, and leucocytes are fixed with formaldehyde. Whole blood samples were labelled with the nuclear dye LDS-751 and with antibodies to HLA-DR or belonging to CD2, CD3, CD4, CD7, CD8, CD10, CD13, CD14, CD19, CD20, CD29, CD33, CD45, CD45RA, CD56, and CD62L (TQ-1) that were directly conjugated to either phycoerythrin (PE) and/or fluorescein isothiocyanate (FITC). Leucocytes were analysed by flow cytometry either in unfixed, unlysed whole blood (15) or after preparation using the Q-Prep system. The binding of eight antibodies, CD19-FITC, CD2-PE, CD3-PE, CD4-PE, CD19-PE, CD29-PE, CD45RA-PE, and CD56-PE, to the surface of lymphocytes was reduced, resulting in significant changes (P F 0.05) in the percentages of cells that stained positively and/or their mean molecules of equivalent fluorochrome (MEF). Further analysis revealed that this was due to the formic acid used during the erythrocyte lysis stage.