Simon, I've done no sorting personally, but I've been in a few discussions on the topic, so here is my quackery: the only way to test for purity of something you sorted is to run it thru flow again. problem 1: your original reagent has been partially quenched in the first pass thru the laser(s). problem 2: if you stain with the same marker but in a different color, the staining will be dim(mer) because of competition for binding sites. my suggestion: do your sorting magnetically or have at least one other marker to verify your purity. example: magnetically sort T cells by CD3, and verify with fluorescent alpha TCR. maciej --- Simon Monard <smonard@trudeauinstitute.org> wrote: > > Hi folks. > Can anyone point me to a discussion of ways to estimate the purity > of a sorted population > of cells. This is clearly easy when you are sorting very bright > cells from negatives > but more problematic when sorting cells from a "shoulder". I > remember seeing a nice > discussion on this subject but cannot remember where. I've been > sorting some very > weakly positive cells from a population, these cells post sort > overlap the negative > cells quite a bit. My customer seems unhappy about that. > Thanks > > Simon Monard > FACS Lab Manager > Trudeau Institute > Saranac Lake > NY12983 > > Ph 518 891 3080 X352 > __________________________________________________ Do You Yahoo!? Make a great connection at Yahoo! Personals. http://personals.yahoo.com
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