Lost cells

From: Keith Kelley (kelley@wh.bayer.com)
Date: Fri Oct 24 1997 - 08:07:32 EST


This is a problem that has no single solution.  In my experience
I have lost cells due to staining/washing, lysis, adherence to various
tube materials, phases of the moon, etc.  There have been any number of
individuals (myself included) who have tried to make an explantation to
some irate individual who put 10,000 cells into 1 ml and wanted to know
why I was not able to give them 10,000 events (sometimes they wanted
10,000 sorted cells, but that is another story).  One approach to take 
is to get some large (cell-sized) fluorescent beads, put 10,000 of them
into the volume you normally use, and run them through the instrument.  
The advantage there is that you have consistent density & fluorescence 
that you can trigger on and eliminate the "no-see-ums" that seem to be
inherent in any biological population.  This should give you a handle on
what % of the population you are losing due to instrumental anomalies
and set a baseline from which to start looking for the disappearing cells
(you can work with any number of beads, as long as you know what it is,
and calculate your % loss from that).

Keith
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