The dye-dilution assay is a powerful tool to study lymphocyte expansion dynamics. By combining time course dye-dilution experiments with computational analysis, quantitative information about cell biological parameters, such as percentage of cells dividing, time of division, and time of death, can be produced. Here, we describe the method to generate quantitative cell biological insights from dye-dilution experiments. We describe experimental methods for generating dye-dilution data with murine lymphocytes and then describe the computational data analysis workflow using a recently developed software package called FlowMax. The aim is to interpret the dye-dilution data quantitatively and objectively, such that cell biological parameters can be reported with an appropriate measure of confidence, which in turn depends on the quality and quantity of available data.
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This work was supported by NIH grant R01 AI132731 (A.H.) and from NIH-NCI CCSG: P30 014195, and the Helmsley Trust (M.N.S.).
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