An indirect rosette technique for the identification and separation of human lymphocyte populations by monoclonal antibodies: a comparison with immunofluorescence methods.
A simple indirect rosette technique is described which allows the rapid enumeration, morphological identification and separation of monoclonal-antibody (MoAb)-defined human lymphocyte populations. The method is based on the binding to MoAb-stained cells of marker ox red blood cells coated with anti-mouse immunoglobulin. When compared with indirect immunofluorescence staining assessed by microscopy, the rosette method is quicker, shows greater sensitivity and is less subjective. Purification of helper (OKT4), suppressor (OKT8) or B-cell (BA1) subpopulations by positive or negative enrichment of rosetted cells gives purity and recoveries comparable with that of the fluorescence-activated cell sorter (FACS). Such purification is readily performed without loss of viability or function and is more easily done sterilely than with the FACS.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000921:Antibody-Producing Cells / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D008214:Lymphocytes / Q000145:classification* / Q000201:enzymology / Q000276:immunology; D051379:Mice; D012397:Rosette Formation / Q000379:methods*; D013601:T-Lymphocytes / Q000201:enzymology / Q000276:immunology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology
D000911:Antibodies, Monoclonal