Hi Cliff, This is my first message to the list which I find a great asset to beginners like me. I thought I'd share a couple of problems we've had. We sort B220+ cells from a number of tissue cell suspensions and attempt ELISPOT assays (measuring frequency of antibody secretors) directly after sorting. We have found that sorting with a BD FACStar plus did not seem to affect frequencies but Mo Flo sorting does (the cells do not secrete). This may be due to the higher pressures involved with Mo Flo sorting. We have tried a number of sheath fluids and sorting at "high" or "low" speeds without success. We have never resolved this problem. Sorting CD8+ T cells and then attempting direct CTL assays afterwards does not seem to be affected. In a nutshell, function of sorted cells can be affected but it is relative to the cell type sorted. I would love to hear any comments by others on this problem. Cheers, Sam ********************** Dr Sam Hou Lung Immunology Group Edward Jenner Institute for Vaccine Research Compton Newbury RG20 7NN web site: www.jenner.ac.uk Ph: 01635-577-924 Fax: 01635-577-901 -----Original Message----- From: Cliff McArthur [mailto:cytocliff@netscape.net] Sent: 06 March 2000 17:17 To: cyto-inbox Subject: HOW viable after high-speed sorting? Hi there, flow aficionados. I know we've all addressed the post-sort viability (after high-speed sorting) question many times over. What I am looking for is not "whether" but "how much," so to speak. To that end, I'd like to ask the List if anyone knows of any published work that has addressed the question not of whether or not cells are viable after such sorting (we know they are, or can be) but "how" viable, that is, do they produce less cytokine or other product, proliferate a little more slowly, or survive for less time, etc., etc. under the same conditions versus "low-speed" sorted cells? Shucks, references that just address this question, if it is not the primary objective of the work, would be great to have. If anyone is willing to share their unpublished insights, I'd love to hear them as well. Thank you very much, Cliff McArthur University of California at San Francisco Departments of Medicine and Immunology/Microbiology 415-502-6860 ____________________________________________________________________ Get your own FREE, personal Netscape WebMail account today at http://webmail.netscape.com.
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