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  • Title: Three-color immunofluorescence analysis of mouse B-lymphocyte subpopulations.
    Author: Parks DR, Hardy RR, Herzenberg LA.
    Journal: Cytometry; 1984 Mar; 5(2):159-68. PubMed ID: 6370630.
    Abstract:
    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.
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