Hi Lynn, as you see from the responses it's a matter of strategy. For sorting of rare cells for subsequent analysis or culture the rarest has been 1 in 500,000. We could have done better than that but the experiment was stopped here. One of the approaches I use to sort rare cells (that cannot be pre-enriched)is to try and increase the no. of possible sort events to enhance recovery and purity. An example; ES cells, less than 1% GFP positive -transient expression, which means I only get the one chance to sort. These cells are grown in the presence of feeder cells, which in turn have a higher autofluorescence and fit rather nicely within the GFP gate; this increases the sort gate to 10% or above (whatever) thereby increasing recovery and purity and decreasing the no. of abort events. So the users are asked not to be too exact about getting rid of the feeder cells (I call them "carrier cells") before sorting. Following this I believe that any cell no matter how rare can be isolated. Ref: E.Paraskeva, A.Atzberger, M.Hentze A translational repression assay procedure (TRAP) for RNA-protein interactions in vivo. PNAS:Vol.95, pp. 951-956, Feb.1998 Regards Ann At 13:39 26.01.01 -0500, you wrote: > >Hello all, > >This question arises from a rather heated discussion following a seminar >yesterday, in which the speaker claimed that flow cytometry is not >useful for analyzing cells that are less than 1-2% of the starting >population. I am sure that with all of the sorting, multicolor analysis, >and multiparameter gating that people do, we can prove this assertion >wrong. > >If you have experience or publications with analysis of events well >below 1% of the starting cell population, could you please share some >examples or references? > >Thanks in advance for your help! >********************************************** >Lynn B. Dustin, Ph.D. >Center for the Study of Hepatitis C >Rockefeller University >Box 64 >1230 York Ave. >New York, New York 10021 >Phone: 212-327-7067 >email: dustinl@mail.rockefeller.edu > >
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