Hi Tony, One way which works well is to perform a proliferative index (PI=S+G2/M) using cell cycle analysis. Cells are permeabilized, fixed and stained with propidium iodide. For simultaneous surface analysis, FITC-conjugated ab are added before permeabolization. Take a look at our paper in Clin Exp Immunol 1996, 104: 412-418 (PT Lukey et al). The Kinetics of DNA histogram (cell cycle analysis) of CD4+ T-cells obtained following antigen stimulation correlated well with results obtained using conventional thymidine uptake. Good luck. Stan ..................................................................... I'd like opinions on the use of flow for the detection of T cell proliferation. I know that bromodeoxyuridine is an established method, but right now out of my reach ($$). My thought is to use anti-CD3, -CD4 and/or -CD8 on bulk lymph node cultures stimulated with or without antigen to get changes in total numbers of cells. Thanks, Tony --
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