PFA fixation of clinical samples

Kunwar, Shaan S_Kunwar at fccc.edu
Mon Feb 13 11:31:41 EST 2006


Hello,

We do routinely fix our clincial samples.  If we must, we ran as soon as 15
minutes after fixing, however we observed an 'incomplete' fixing of certain
populations, leading to larger/dual clusters.  

We found overnight in 1% PFA yields uniform populations, I would think two
hours would suffice the remodeling as well as neutralize the biohazard(s).


When I arrived here, the lab was in transition from live to fix.  It's
adoption was simply to provide a break point, at which clinical samples may
be preserved overnight.  It required validation and review by the then
supervisor and director.  Once it was determined that the signal was not
diminished, it was incoorperated.  The cells appeared smaller on scatter,
our CVs were reduced as well.  

The FCM lab here routinely process marrows, bloods, tissues, FNAs, and more.


Shaan Kunwar MS
Fox Chase Cancer Center
Protocol Support Laboratory


-----Original Message-----
From: Jonni S. Moore
To: cyto-inbox
Sent: 2/9/2006 9:59 AM
Subject: PFA fixation of clinical samples

Hi Clinical Folks,

I have a quick question...do you routinely PFA fix your bone marrows and
bloods and if so how long do you wait to run them after fixation.  Old
dogma held that you should not run anything for at least 2 hrs after
fixation until the "remodeling " that occurs is complete.  If you do not
fix (which for analysis purposes is preferable as far as I am concern),
how do you deal with biohazard concerns?  Thanks for any comments!

Cheers
Jonni
-- 
Jonni S. Moore, Ph.D.
Associate Professor of Pathology and Laboratory Medicine
Director, PATH BioRESOURCE
Director, Flow Cytometry and Cell Sorting Shared Resource, Abramson
Cancer Center
Director, Clinical Flow Cytometry, Hospital of the University of
Pennsylvania
University of Pennsylvania School of Medicine
220 John Morgan Bldg.
Philadelphia, PA  19104-6082
Phone:	215-898-6853
FAX:	215-898-0734



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