Platelet Activation Studies in Pheresis

April G. Durett/MDACC April_G._Durett/MDACC%MDACC at notes.mdacc.tmc.edu
Mon Feb 10 08:56:18 EST 1997


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 at notes.mdacc.tmc.edu@internet





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