Absolute counts from flow cytometry

Dennis_Young@CIS.ucsd.edu Dennis_Young at CIS.ucsd.edu
Fri Nov 15 17:21:00 EST 1996

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 J. Young                            Voice : (619) 822-0407     *
* Flow Cytometry Core Facility               FAX   : (619) 822-0412     *
* University of California, San Diego  USA   e-mail: djyoung at ucsd.edu   *
*                       http://www-core.ucsd.edu/flow-core.html         *
______________________________ Reply Separator _________________________________
Subject: Re: Absolute counts from flow cytometry
Author:  douglas at 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


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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