Quantitative analysis of calcium (Ca2+) mobilization after stimulation with mitogens or anti-CD3 antibodies. Simultaneous fluo-3 and immunofluorescence flow cytometry.
A method is described for flow microfluorometric analysis of calcium mobilization in immune cells. This method is based on dual-color analysis of heterogeneous cell populations using fluo-3 and phycoerythrin (PE)-conjugated monoclonal antibody (mAb). This technique allows the detection of fluo-3 fluorescence as a measure of an increase in cytoplasmic free calcium ([Ca2+]i) while simultaneously discriminating PE-mAb-labeled cells without using wavelength ratio imaging. This method provides a unique way to distinguish patterns of calcium mobilization within a heterogeneous cell population.
Sei
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Laboratory of Neuroscience, NIDDK, National Institutes of Health, Bethesda, MD 20892.
Arora
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PK
D000814:Aniline Compounds; D000818:Animals; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000276:immunology*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology; D001323:Autoantibodies / Q000494:pharmacology; D017252:CD3 Complex; D015704:CD4 Antigens / Q000276:immunology; D016827:CD8 Antigens; D002118:Calcium / Q000032:analysis*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D008213:Lymphocyte Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D008934:Mitogens / Q000494:pharmacology; D010799:Phycoerythrin; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D018800:Thy-1 Antigens; D014966:Xanthenes
D000814:Aniline Compounds; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D000954:Antigens, Surface; D001323:Autoantibodies; D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; D008934:Mitogens; D011948:Receptors, Antigen, T-Cell; D018800:Thy-1 Antigens; D014966:Xanthenes; D010799:Phycoerythrin; C059715:Fluo-3; D002118:Calcium