Analysis of cell death by apoptosis is illustrated here by the uptake of Hoechst 33342 and exclusion of PI in a method described by Omerod et al, and Dive, et al. Reportedly, a short exposure of apoptotic cells to HO33342 labels these cells more brightly , perhaps through increased membrane permeability. Viable, nonapoptotic cells , require a much longer exposure to obtain comparable fluorescence intensity. Thus, by the supravital uptake of HO33342 combined with PI exclusion, and analysis of light scattering characteristics of the cells, multiple populations of cells can be identified. In this experiment, mouse lymphocytes from animals treated in-vivo with IL-2 and anti-CD3 antibody are studied.
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