Re[2]: Absolute counts from flow cytometry

Dennis_Young@CIS.ucsd.edu
Fri, 15 Nov 1996 14:21:00 -0800

Be careful. I tried this protocol cited below and you see a lot of debris also
stains. Not very satisfactory dead/live discrimination either.

Use calibrated beads: add a known concentration of beads to all samples and
calculate the ratio of WBC:Beads. (BDIS has a product called TrueCOUNT - tubes
with beads.

Dennis
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______________________________ Reply Separator _________________________________
Subject: Re: Absolute counts from flow cytometry
Author: douglas@HELIX.NIH.GOV at @UCSD
Date: 11/15/96 8:38 AM

At 04:29 PM 11/14/96 EST, you wrote:
>
>We are trying to generate absolute counts for different cell types in the
>peripheral blood of mice. What are your preferred methods? ......

Jeff Carrell
Human Genome Sciences

Jeff,

You may want to look at LDS-751. It is a nucleic acid dye not requiring
permeabilization and can be used in whole blood with no lysing.
See the following reference:

Multidimensional Flow Cytometric Blood Cell Differentiation without
erythrocyte lysing, Terstappen, et al. in Blood Cells (1991) 17:585-602.

The dye is available from Exciton, Inc (513) 258-3937, Dayton, Ohio.

Good luck.

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>Subject: Re: Absolute counts from flow cytometry