counting without an isotype

Fredda London flon at temple.edu
Fri Oct 21 09:25:25 EST 2005


Hi Judith,
If you truly wish to set a region for microparticles, spin down your 
cells at a mild rate that will not lyse them.  Then spin the 
supernatant harder to remove smaller cells.  Then either run the super 
through analysis, or (better) high speed spin your supernatant 100,000 
xg for 1-4 hours and resuspend pellet in buffer to analyze and set a 
positive window.  This will differentiate between cellular 
microparticles and aggregates of label and debris that confuse the 
issue.

Hope that is helpful.
Fredda London


On Oct 20, 2005, at 6:51 AM, Stewart, Judith wrote:

> fellow flowers
>    I have no problem giving up the isotype for my surface marker work, 
> we
> use the MFI almost exclusively now.  What I am puzzled by is how to 
> count
> particles (microparticles to be specific)? Do I just just my FMO to 
> set the
> boundary and count only those above that level or do I take the total 
> count
> and subtract the count from the FMO tube?  Many thanks.
>
Fredda London, Ph.D.
Associate Professor
Sol Sherry Thrombosis Research Center
Temple University School of Medicine
3400 North Broad Street
Philadelphia, PA 19140
phone: 215-707-4458
fax: 215-707-3005




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