Too true, also helps to watch the abort rate especially with bigger/stickier cells. I use 30psi, 100um nozzle mostly but the cell concentration depends on the cell type. Ann Atzberger David.C.McFarland@gsk.com wrote: Janet, There is no one best concentration. You can make some educated guesses but sometimes you have to determine this empirically. There are several considerations including cell size, nozzle size, sheath pressure, sorting electronics and sample stickiness. For example, when doing high-speed (30,000/sec), high-pressure (45 psi) sorting with a 70 micron nozzle on a digital Vantage, the most important factor in my hands is the sample. For PBMCs, I request cells at a concentration of 50 to 100 million/mL and I filter through 50 micron mesh immediately before placing on the instrument. But this setup would not work for an adherent cell line that clumps badly and/or that contains much larger cells. Contact me directly if you would like to go into more specific details about your particular issues and I will try to help. Dave David McFarland GlaxoSmithKline ----- Forwarded by David C McFarland/PharmRD/GSK on 25-Aug-2004 09:14 AM ----- "janet dow" <jldow@unity.ncsu.edu> 24-Aug-2004 12:09 To "Cytometry Mailing List" <cytometry@flowcyt.cyto.purdue.edu> cc Subject optimum concentration for sorting Dear Flowers: What do people find as the optimum concentration of their samples for sorting? I am speaking in terms of the best concentration for keeping the machine happy during a sort and keeping the drop delay constant. Thanks in advance for all your assistance. Janet Dow -- Janet Dow Laboratory Research Specialist and Manager Flow Cytometry and Cell Sorting Facility North Carolina State College of Veterinary Medicine 4700 Hillsborough Street Room C-309 Raleigh, NC 27606 (919)513-6443 --------------------------------- ALL-NEW Yahoo! Messenger - all new features - even more fun!Received on Thu Aug 26 16:18:00 2004
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