There are an array of problems in using a cell sorter to sort "rare" events. This is what we thought to be possible many years ago in using a sorter to sort stem cells for transplantation. The reality is a numbers game. Sorting through 10^9 cells takes a long time even at warp sort speed. 10^9 starting populations is probably on the low side. The more efficient way to look for these cells is by depletion of known unwanted cells prior to sorting. This can be done easily with magnetic columns. The days of using a cell sorter to sort for hours and hours should come to an end. I have seen 10^12 cells applied to a Clinimacs Column and depleted of a 65% population of T cells to virtually none remaining in a matter of hours. Never could do this with a sorter. For this project I would try to either deplete the population of unwanted cells then sort or enrich for the 1/10^6 cells using a clinimacs or automacs, then sort if needed to further define the population. The second way would be far cheaper. Sort time would most likely be less than 1 hour. Jim Houston Coordinator Flow Cytometry and Cell Sorting Stem Cell Transplantation St. Jude Children's Research Hospital > ---------- > From: Todd Belanger > Sent: Monday, August 12, 2002 9:23 AM > To: Cytometry Mailing List > Subject: sorting rare events- 1 in a million or more > > Hi, > > I am new to sorting (but I have ten years of flow experience) and we just > purchased a FACSVantage/DiVa. Some of our projects require sorting rare > cells at levels of 1 in a million or ten million. Some of the researchers > say it could be one in 100 million (which seems quite impossible to me). > Does anyone have any pointers or particularly good papers that would help > me in this task? How low can you go (in terms of rare events) and still be > relatively confident in what you sorted? Currently the researchers I will > be doing the sort for has two markers- PI to discriminate live cells and a > FITC conjugated marker. I know more markers would be better for > discriminating rare events but their doesn't seem to be any for this > particular experiment. > > Any and all help would be greatly appreciated. > Todd > > Todd J. Belanger > Lab Manager > Cellular Immunology > Vaccinex, Inc. > 1895 Mt. Hope Ave. > Rochester, NY 14620 > email: tbelanger@vaccinex.com > www.vaccinex.com > >
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