Looking for tips/feedback on doing a 3-color analysis of platelet activation to compare degranulation (BD: CD62-PE) and gpIIb/IIIa activation(BD: PAC-1-FITC) in heparinized human whole blood also labeled with CD61 (BD: CD61-PerCP)as the pan-platelet marker. Ultimately the purpose is to examine/compare platelet (and WBC) activation events upon exposure of blood to common and surface-modified versions of medical plastics used in cardiopulmonary bypass applications. Any suggestions on optimal buffers and whether or not to use fixatives? I'm somewhat new in the area but my limited experience tells me that care is required in choice of the latter to avoid cell lysis. M. Wolf
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