Single step isolation of immunofluorescence-stained, viable lymphocytes by fluorescence-activated cell sorter using ethidium bromide.
The paper presents a method for preparation of viable, immunofluorescence-stained lymphocytes in a single isolation step by fluorescence-activated cell sorter. Discrimination between viable, immunofluorescence positive cells and nonspecifically stained dead cells was carried out on the basis of the DNA-specific dye ethidium bromide. Ethidium bromide enters cells with damaged plasma membrane only. The method is described by the example of isolation of Lyt-2 positive mouse lymph node cells.
D000818:Animals; D004996:Ethidium; D005434:Flow Cytometry / Q000379:methods; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D008198:Lymph Nodes / Q000166:cytology; D008214:Lymphocytes / Q000166:cytology*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D013861:Thiocyanates
D005452:Fluoresceins; D005456:Fluorescent Dyes; D013861:Thiocyanates; D004996:Ethidium; D016650:Fluorescein-5-isothiocyanate