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.