Analysing cell division in vivo and in vitro using flow cytometric measurement of CFSE dye dilution

Since its introduction in 1994 (J. Immunol. Methods 171 (1994) 131), the fl ow cytometric analysis of lymphocyte proliferation by serial halving of the fluorescence intensity of the vital dye CFSE (carboxyfluorescein diacetate , succinimidyl ester or CFDA-SE) has become widely used in immunological la boratories around the world. This technique allows the visualisation of eig ht to 10 discrete cycles of cell division by Row cytometry, both in vitro a nd in vivo. Appropriately conjugated antibodies can be used to probe surfac e marker changes as cells divide, or changes in expression of internal mole cules such as cytokines when appropriate fixation and permeabilisation prot ocols are used. An added advantage of the technique is the ability to recov er viable cells which have undergone defined numbers of cell divisions by f low cytometric sorting, allowing functional studies to be performed. Other commonly used assays of cell proliferation give only limited information, a s they usually measure division at a population level. The CFSE technique c an be used to determine kinetics of immune responses, track proliferation i n minor subsets of cells and follow the acquisition of differentiation mark ers or internal proteins linked to cell division. (C) 2000 Elsevier Science B.V. All rights reserved.