title: Tips and tricks for flow cytometry-based analysis and counting of microparticles.
abstract: 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.
authors.affiliation: Research & Technology Department, BioCytex, Marseille, France. Electronic address: philippe.poncelet@biocytex.fr.
authors.forename: Philippe
authors.initials: P
authors.lastname: Poncelet
authors.affiliation: VRCM, UMR-S1076, Aix-Marseille Université, INSERM, Faculté de Pharmacie, Marseille, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Robert
authors.affiliation: Hematology Laboratory, Namur Thrombosis and Hemostasis Center (NTHC), Namur Research Institute for Life Sciences (NARILIS), CHU UCL Namur, Université catholique de Louvain, Yvoir, Belgium.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bailly
authors.affiliation: INSERM U1098, EFS BFC, Université de Bourgogne Franche-Comté, Besançon, France.
authors.forename: Francine
authors.initials: F
authors.lastname: Garnache-Ottou
authors.affiliation: Research & Technology Department, BioCytex, Marseille, France.
authors.forename: Tarik
authors.initials: T
authors.lastname: Bouriche
authors.affiliation: Department of Hematology, Namur Thrombosis and Hemostasis Center (NTHC), Namur Research Institute for Life Sciences (NARILIS), CHU UCL Namur, Yvoir, Belgium.
authors.forename: Bérangère
authors.initials: B
authors.lastname: Devalet
authors.affiliation: International Consultancy in Blood Component Quality/Safety Improvement, Audit/Inspection and DDR Strategy, London, UK.
authors.forename: Jerard H
authors.initials: JH
authors.lastname: Segatchian
authors.affiliation: INSERM U1098, EFS BFC, Université de Bourgogne Franche-Comté, Besançon, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Saas
authors.affiliation: Hematology Laboratory, Namur Thrombosis and Hemostasis Center (NTHC), Namur Research Institute for Life Sciences (NARILIS), CHU UCL Namur, Université catholique de Louvain, Yvoir, Belgium.
authors.forename: François
authors.initials: F
authors.lastname: Mullier
mesh_terms: D000818:Animals; D055252:Cell-Derived Microparticles / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans
keywords: Flow cytometry; Labile blood products; Microparticles; Standardization; Transfusion