Right angle light scatter: a necessary parameter in flow cytofluorimetric analysis of human peripheral blood mononuclear cells

The techniques of flow cytometry are being increasingly used to classify lymphocytes according to their surface antigens. Accurate analyses depend on the exclusion of non-lymphocytic contaminant cells. Using a fluorescence-activated cell sorter (FACS IV), virtually complete separation of human lymphocytes from monocytes and granulocytes can be achieved by means of wide-angle (90°) light scatter. The technique described is simple and suitable for routine use. Used in conjunction with fluorescein-labelled monoclonal antibodies and logarithmic amplification of the fluorescence signals, it improves the accuracy of lymphocyte subset analysis. Its use is particularly necessary in the study of subjects such as renal allograft recipients, who have a high or variable contaminant cell:lymphocyte ratio.