Positive selection of mouse B and T lymphocytes and analysis of isolated populations by flow cytometry.
Mouse spleen cells were separated into Ig+ and Ig- populations by positive selection using petri plates coated with rabbit anti-mouse immunoglobulin. The Ig- cells were subsequently incubated with mouse monoclonal alloantisera to Thy1.2 prior to a second positive selection. The adherent populations were characterized as B (Ig+) or T (Thy1.2+) lymphocytes on the basis of surface immunofluorescence and mitogen-induced proliferation. Analysis of the 2 isolated populations by flow cytometry showed that B and T lymphocytes could be distinguished by their forward light scatter as well as their fluorescence after incubation with fluorescein diacetate.
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D001402:B-Lymphocytes / Q000276:immunology*; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000032:analysis; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000201:enzymology; D008297:Male; D051379:Mice; D011947:Receptors, Antigen, B-Cell / Q000032:analysis; D013154:Spleen / Q000166:cytology; D013601:T-Lymphocytes / Q000276:immunology*; D018800:Thy-1 Antigens
D000954:Antigens, Surface; D011947:Receptors, Antigen, B-Cell; D018800:Thy-1 Antigens; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase