Sorting woes

From: Steve G. Hilliard (steve@habanero.cb.uga.edu)
Date: Thu Aug 17 2000 - 16:25:35 EST


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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