LSR help needed

From: bunny (bunny@cotleur.com)
Date: Fri Nov 09 2001 - 00:07:47 EST


We recently acquired a BD LSR. Switching over from a Facscan
is a bit like patting your head and rubbing your stomach- in
the opposite directions.  The visual cues are a bit off from
the usual Fascan display, and I'm having trouble getting my bearings.

Running your standard lysed whole blood prep, I find it
practically impossible to achieve anything close to the
"Facscan traditional" 3-part Dif display. At the most I can
get Lymphs & monos, but no grans. If I manage to have all
three in the view- then the monos overlap the lymphs and
cannot be gated separately.
More disturbing (to me) is that I cannot get the lymphs off
the SSC low axis. I've pumped up the PMT voltage, then tried
toggling throught the 2-4-8 log scale. All I've managed to
so was increase the spread between the populations, or smear
out the lymphs- but the population itself will NOT move up
the axis. What am I doing wrong?


I'm also having a heck of a time comping the APC from the
FL3 channel (using FlowJo). All the other parameters look
fine, this one won't budge. If anyone has any special
tips-please pass them on!


Bunny Cotleur
Cleveland Clinic Foundation
Neurosciences   NC30
9500 Euclid Avenue

Cleveland, OH  44195

(216) 444-1164
cotleua@ccf.org



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