Hi, I thought that since there was so much interest in CFSE I would share my recent experiences. I hope it will be of some use. I have been labelling human PBMC with CFSE according to Current Protocols methodology. After labelling, my cells were more than 95% viable by trypan blue exclusion... but would not proliferate in culture. In the end, by titration, I found that reducing the final concentration of CFSE to 2 micromolar, (instead of the suggested 10 micromolar) gave good staining while still allowing good proliferation. So the moral of the story seems to be that CSFE can do funny things to cells, and although they are viable they may not be fully functional! -- Lisa M Gale Department of Microbiology & Immunology University of Adelaide ph +61 8 8303 4630 fax +61 8 8303 4362 email lisa.gale@adelaide.edu.au
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