Dear Flowers, When doing transfection experiments, we routinely determine viability of the samples by FACS on the basis of FCS vs SSC profiles (these are non-adherent cell lines). Someone in the lab has just done an experiment where they checked the viability of each sample by trypan blue exclusion by light microscopy as well as by FACS. To her surprise viabilities were much higher by trypan blue than by FACS (eg 60% vs 17%, respectively). One way to explain this is that maybe non-viable cells break up so one single non-viable cells becomes several "events" in the non-viable area of a FACS plot, leading to a disproportionate number of non-viable vs viable events. Does anyone else have any suggestions, or seen this disceprancy themselves? Which is more accurate? I would always have said FACS but now I'm not so sure. Thanks in advance, Michelle ------------------------------------------------ Michelle Miller JJR Labs GPO Box 3331 Sydney NSW 2001 Australia 02 360 9377 (Ph) 02 360 9813 (fax) jjph13mm@angis.su.oz.au
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