Detection of Platelet Activation by Flow Cytometry.

Phycoerytherin-labeled antibody to CD62P (P-selectin) (Becton Dickinson, San Jose, CA) at a dilution of 1:100 was added to platelets after stimulation or drug treatment of 30 minutes. Platelets were then fixed in 1% formalin. Surface P-selectin was measured by flow cytometry (LSRII; Becton Dickinson). To detect conformational changes in GPIIbIIIa, fluorescein isothiocyanate (FITC)-fibrinogen (Abcam) was added for 30 minutes, and platelets were analyzed by flow cytometry. We have previously used these techniques to measure platelet activation (Zhu et al., 2014).