Quantitative fluorescence flow cytometry: a comparison of the three techniques for direct and indirect immunofluorescence.

摘要: Three types of microbead calibrators available for quantitative fluorescence flow cytometry have been studied in parallel using a variety monoclonal antibodies (MoAbs). The QIFI kit is designed indirect immunofluorescence (IF), and both the Quantum Simply Cellular (QSC) assay Quanti-BRITE are direct IF. Because different nature respective ligands, epitopes on cells versus F'ab-portions QSC beads, large differences titration curves number CD MoAbs were noted between beads cells. Use fluorescein isothiocyanate (FITC) phycoerythrin (PE) conjugates same reagent revealed substantially numbers cellular binding sites. Numbers sites as determined by IF similar. We also found that erythrocyte (RBC)-lysing reagents cause varying sometimes substantial reduction intensity (FI) stained directly with CD34 MoAb conjugates, but RBC-lysing had no effect FI indirectly MoAbs. This report defines variables critical standardized cytometry. conclude choice calibrators, fluorochrome staining methods, modes sample processing can determination to Direct appear yield comparable results