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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