On 1/13/04 10:53 AM, "Polakoff, Dixie" <DPolakoff@pdl.com> wrote: > I have been asked to post this question for one of my investigators: > > I would like to know what speed (events/second) is ideal to sort CD4/CD45Rbhi > mouse T > cells for adoptive induction of inflmmatory Bowel disease in scid mice. > I am looking at several parameters that might have an effect on disease > induction and > speed sort could be one of the parameters > > Dixie Polakoff > Protein Design Labs, Inc. > Please provide additional information for the group. What machine are you using, and at what pressure and frequency? I'll try to answer without this info: In terms of "ideal speed" for colitis, theoretically the speed should not matter. As long as the cells to be injected are alive and the post-sort population has the profile of interest, the speed should not be relevant (within reason). Over the years, I have done this particular sort more times than I'd care to count, in the hundreds for sure. Years back on a FACStar or EPICS Elite ESP, I'd typically sort at relatively low PSI and speed. Today on our DiVa, I use a 70 micron nozzle, and a still relatively conservative PSI, often times around 27 psi. If you use the jet parameter optimizer at Scripps' web site, http://facs.scripps.edu/optimize3.html (thanks guys, credit where credit is due), you'll end up with a frequency around 61,000. For this particular sort, I'll often use a speed in the neighborhood of 9,000. However, I've done it faster when significantly large numbers of cells were needed for the experiment. If you have any specific questions, feel free to ask. -- Samuel Connell Imaging Facility and Technology Manager La Jolla Institute for Allergy and Immunology 858-558-3508 sconnell@liai.orgReceived on Fri Jan 16 14:38:00 2004
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