Detection Of Platelet-derived Microparticles By Flow Cytometry

Objective This study is to establish optimization of flow cytometry (FCM)method for quantification of platelet- derived microparticles(PMPs) and systemically evaluated.Method platelet rich plasma (PRP)was prepared as samples, and then were stained with a fluorochrome-conjugated monoclonal antibody directed towards platelet surface glycoprotein. 0.82μm latex beads was used to calibrate of forward scatter threshold parameter and setup optimization of procedure of flow cytometry and as standard for counting PMPs. PMPs generated in vitro using ADP or collagen were detected as positive control, and systemically evaluated the method.Results 30 healthy donors was detected ,the resting PMPs was 2.19%±0.48%, after activated with ADP, the PMPs was 5.43%±3.48%, when stimulated with collagen the PMPs was 3.46%±1.14%. Compared with resting PMPs, the activated PMPs notably increased. Samples collected into two different anticoagulant 109mmol/L acid-citrate-dextrose(ACD) and CTAD(ACD0.11mol/L,Theophylline15mmol/L,ERI AAF 3.7mmol/L,Dipyridamole0.198mmol/L). The resting PMPs of them were remarkably different. The storage time of sample impacted the results of PMPs. 37 patients with different thrombus disease was detected, the PMPs were evidently increased.Conclusion The method of PRP-FCM to detected PMPs is feasible, and the method is simple, influence factor relatively less, it can be as routine test. PMP is a useful parameter for monitoring platelet activation and in diagnosing thrombus disease early.