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

Fiche publication

Date publication

octobre 2015

Journal

Transfusion and apheresis science : official journal of the World Apheresis Association : official journal of the European Society for Haemapheresis

Auteurs

Membres identifiés du Cancéropôle Est :
Pr GARNACHE-OTTOU Francine

Tous les auteurs :
Poncelet P, Robert S, Bailly N, Garnache-Ottou F, Bouriche T, Devalet B, Segatchian JH, Saas P, Mullier F

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/26603057

Résumé

Submicron-sized extra-cellular vesicles generated by budding from the external cell membranes, 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.