Hi folks, I just attempted a sort for someone, and had a problem I've never had before. The instrument's a 753, w/ a 75um orifice, operated at the usual parameters. Sort setup went fine, I loaded the sample, set the sort gate, and started the sort. I was also running our other instrument, so I stepped out to start those samples, but when I checked in on the sorter droplet formation had ceased and the plates were wet. I stopped, cleaned, futzed, and re-established setup, but the problem would reoccur in a minute or two, making sorting impossible. These are fibroblasts in normal DMEM (I think), w/ 1XPBS sheath--does it sound like I'm getting clogs? The stream still flows, so the flow cell orifice isn't clogged, but something happens that wipes out droplet formation, and it occurs a minute or two after the sample is loaded. I don't have the problem w/ beads, no matter how long they run, so I don't think it's a funky bimorph or unstable electronics. Does this sound familiar to anyone? Any suggestions for troubleshooting? I seem to get very rapid settling with these cells--has anyone used higher viscosity to keep them suspended? I tried raising the sample takeup tube off the bottom and running them slowly, but it made no difference. Thanks in advance, Steve +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Steve G. Hilliard (706) 542-9474 University of Georgia Cell Analysis Facility flowman@uga.edu http://floweb.cb.uga.edu/
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