Three-color immunofluorescence analysis of mouse B-lymphocyte subpopulations.
We have modified a fluorescence-activated cell sorter (FACS) to make three independent immunofluorescence measurements on each cell and used this system to study mouse B-lymphocyte subpopulations. An argon-ion laser (emitting at 488 nm) excites fluorescein- and phycoerythrin-labeled reagents, and a tunable dye laser charged with rhodamine 6G (emitting at 615 nm) excites an allophycocyanin-labeled reagent. We report simultaneous measurements of IgM, IgD, and the recently-defined mouse B lymphocyte antigens BLA-1 and BLA-2 on splenic lymphocytes of CBA/J mice and mice of the congenic strain CBA/N (which have an X-linked immunodeficiency [xid]). These data provide information on relationships among the B-cell populations in CBA/J "normal" mice and the defective CBA/N that could not be derived from one- or two-color immunofluorescent measurements. We believe this is the first use of allophycocyanin as an immunofluorescence label.
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis*; D001402:B-Lymphocytes / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D007072:Immunoglobulin D / Q000032:analysis*; D007075:Immunoglobulin M / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C; D008811:Mice, Inbred DBA; D013045:Species Specificity
D000954:Antigens, Surface; D005456:Fluorescent Dyes; D007072:Immunoglobulin D; D007075:Immunoglobulin M