>Does anyone have experience using CSFE (carboxy fluoroscein succinimidyl >ester sold by Molecular Probes) for staining human primary T lymphocytes. I >have seen it used for staining mouse T lymphocytes (and B lymphocytes) so >that one can follow their replicative history, but have heard through the >grapevine that when human cells divide, they lose all of the CSFE dye. David, Not true; the dye works in human cells just like in mouse. The only way the cells lose all the dye is if they indeed undergo many (>8-10) cell divisions before you assess them by flow. We have used CFSE for human cells using the same protocol as used for mouse with essentially the same technical results. Some people have claimed that labeling at 37C works better for human cells, whereas room temp or 4C works best with mouse. I just label everything at rm temp for about 3 min. AW Andrew D. Wells, Ph.D. University of Pennsylvania Department of Medicine 904 Stellar-Chance Laboratories 422 Curie Boulevard Philadelphia, PA 19104 (215) 898-1951 (215) 573-2880 (FAX) adwells@mail.med.upenn.edu
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