[Cytometry] Fixation of platelets

Nick Greco ngreco at clevelandcordblood.org
Wed Jul 15 11:20:34 EDT 2009


Hello Jim Kurtz,

A direct comparison of fixed (1 or 4% paraformaldehyde prepared from solid
paraformaldehyde versus a 37% solution of formalin which contains 10-15%
methanol) versus non-fixed platelets will likely demonstrate higher binding
of antibodies to the PF-fixed platelets with newly exposed cryptic
non-specific antigen sites.  Using the liquid formalin will likely result in
even higher non-specific binding due to the permeabilization and the
labeling of intracellular structures.

Nick



Nicholas J. Greco, Ph.D.
Cord Blood Bank Processing Facility Director
Cleveland Cord Blood Center
Suite 150
25001 Emery Road
Cleveland, OH 44128-0804
216-896-0384 (office)
216-896-0320 (fax)
440-488-9205 (cell)




Instructor of Medicine
School of Medicine, Division of Hematology/Oncology
Case Western Reserve University
Wolstein Research Building 2-101
2301 Cornell Road
Cleveland OH 44106
216-368-5299/5308
njg7 at case.edu/nicholas.greco at case.edu


-----Original Message-----
From: cytometry-bounces at lists.purdue.edu
[mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of
KurtzJ at usa.redcross.org
Sent: Tuesday, July 14, 2009 3:21 PM
To: cytometry at lists.purdue.edu
Subject: [Cytometry] Fixation of platelets

Hi everyone,

I am interested in anyone that has experience in fixing platelets.  My
main question is that if you are going to store fixed platelets for
10-14 days is it better

to keep the samples at 4oC or at room temperature, or does it make no
difference.  My staining would be done after platelets had been fixed
and stored.

Some people stain and then fix platelets and store them in the cold,
does keeping them in the cold comparing to room temperature storage have
something 

to do with prior staining of the samples before fixing?  Any information
would be appreciated.   Thanks,  Jim Kurtz

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