Re: Platelet Activation Studies in Pheresis

April G. Durett/MDACC (April_G._Durett/MDACC%MDACC@notes.mdacc.tmc.edu)
10 Feb 97 9:56:18 EDT

I have used the BD Procedure : Flow Cytometric Analysis of Platelets; in
addition to the references sited in the procedure I found two other references
to be of assistance:
Transfusion 30:20-25,1990 R Funheer, et al, Detection of Platelet Activation
with Monoclonal Antibodies and Flow Cytometry
Thrombosis & Hemostasis 65:467-473,1991 C Abrams & S Shattil, Immunological
Detection of Activated Platelets in Clinical Disorders

1. Collect the peripheral blood samples in ACD (1:9 ratio) to eliminate any
aggreagation. If you are not using ACD in the pheresis procedure, I would
suggest adding to the pheresis collects also. During venipuncture, use as
large bore needle as possible and withdraw the sample with a 'slow and gentle
touch' so as not to activate the platelets during collection...I would not
suggest using any vacutainer tubes for sample collection unless you add the
sample after releasing the vacuum.
2. During the acquisition, if you want to look at stricly the platelets, set
both the FSC and SSC on log scale and the platelets will be displayed along
the diagonal in the center of the screen, with the RBC in the upper right
corner.
3. I would suggest including a monocyte/granulocyte marker with the platelet
activation markers to distinguish platelets from platelets that are attached to
mono/grans. A double check would be to prepare a slide of the sample left from
acquistion to morphological evaluate for any aggregates or platelets attached
to cells.

April G. Durett
Technology Transfer - Flow Cytometry
Blood & Marrow Transplantation
Univ TX MD Anderson Cancer Center
phone (713)792-4367
e-mail : adurett@notes.mdacc.tmc.edu@internet


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