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.