Tips and tricks for flow cytometry-based analysis and counting of microparticles

作者：Poncelet Philippe; Robert Stephane; Bailly Nicolas; Garnache Ottou Francine; Bouriche Tank; Devalet Berangere; Segatchian Jerard H; Saas Philippe; Mullier Francois

来源：Transfusion and Apheresis Science, 2015, 53(2): 110-126.

DOI：10.1016/j.transci.2015.10.008

• Microparticles
• Flow cytometry
• Transfusion
• Labile blood products
• Standardization

Submicron-sized extra-cellular vesicles generated by budding from the external cellmembranes, microparticles (MPs) are important actors in transfusion as well as in other medical specialties. After briefly positioning their role in the characterization of labile blood products, this technically oriented chapter aims to review practical points that need to be considered when trying to use flow cytometry for the analysis, characterization and absolute counting of MP subsets. Subjects of active discussions relative to instrumentation will include the choice of the trigger parameter, possible standardization approaches requiring instrument quality-control, origin and control of non-specific background and of coincidence artifacts, choice of the type of electronic signals, optimal sheath fluid and sample speed. Questions related to reagents will cover target antigens and receptors, multi-color reagents, negative controls, enumeration of MPs and limiting artifacts due to unexpected (micro-) coagulation of plasma samples. Newly detected problems are generating innovative solutions and flow cytometry will continue to remain the technology of choice for the analysis of MPs, in the domain of transfusion as well as in many diverse specialties.

• 出版日期2015-10