RE: cell count

From: Carl-Magnus Hogerkorp (carl-magnus.hogerkorp@molmed.lu.se)
Date: Wed Dec 08 1999 - 03:54:57 EST


Dear Chris,

Provided that the preparation is OK ,the key is off course to evaluate a
correct drop delay value ,which we do every day running a drop delay profile
with beads even if it only differs at most .1 of a drop from one run to the
other given that other settings all the same ( i.e. pressure, frequencies,
amplitude etc.). Variations may occur, depending on the nozzle used, how
clean it is and variations in sheath pressure, this is at least my
experience with the Vantage SE which has a fluidics system more sensitive
than the Vantage and probably the Star Plus too. For example I recently
experienced a sudden drop from a normal 95-100% to >70 which probably was
due to a divergence in the sheath pressure as a result of modifications of
the sheath tank.

Carl-Magnus Hogerkorp
Stem Cell Lab.
University Hospital of Lund
Sweden

-----Original Message-----
From:	Marty Bigos [mailto:bigos@stanford.edu]
Sent:	den 3 december 1999 20:20
To:	Cytometry Mailing List
Subject:	Re: cell count


We have had users getting recoveries significantly different from the
Facstar counter as well. Although, at times, this could depend on the
sample prep and sort conditions, a test run in parallel between a
group that got consistently good agreement and a group which had low
(< 80%) agreement revealed problems with the method of handling and
counting the cells in the collection tube. Coating the collection
tube with serum to prevent cells sticking, spinning the cells down
harder, and using the appropriate cover slip in the hemocytometer
resulted in the counts of the collection tube being within 10% of the
counts on our Facstar.

Marty Bigos
Stanford Shared FACS Facility

>Hello gang,
>My question deals with our FacStar. I am getting reports of counts that are
>about 1/2 of the cell number the instrument says were recovered. I am
>wondering if anyone has seen this or what to do to fix this problem. Cell
>count is very important in many of our studies.
>Thanks in advance and Happy Holidays,
>
>chris
>
>
>
>
>
>Chris Lena
>Laboratory Technician
>La Jolla Institute For Allergy and Immunology
>phone:(858)678-4536
>fax:(858)678-4595
>cali@liai.org
>http://www.liai.org
>
>               _   _
>              (_)-(_)
>                \"/
>                =V=
>
>



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