title: Light-scattering gating and characterization of plasma microparticles.
abstract: Flow cytometry method (FCM) is widely used for analysis of cell-derived microparticles (MPs). Numerous efforts are currently aimed to standardize these measurements among different instruments. We push the FCM characterization of MPs to the limit based on rigorous simulation of measured signals. We measured forward- and side-scatter (FSC/SSC) signals and angle-resolved light-scattering profiles (LSPs) of polystyrene microspheres and MPs, including their aggregates, using a scanning flow cytometer (SFC). We used the Mie theory to (1) accurately evaluate instrument detection limits; (2) construct FSC/SSC gates for MPs in absolute scales of size and refractive index (RI); and (3) determine size and RI of individual spherical MPs. LSPs were used for advanced characterization, including differentiation of spherical and nonspherical particles. The proposed absolute FSC/SSC gating is naturally standardized for any FCM instrument, given the knowledge of its optical system and leads to instrument-independent analysis of MPs. The inverse Mie problem has a unique solution only for some regions of size and RI and uncertainties rapidly increase with decreasing size and RI. The developed methods are applicable to any flow cytometer, but are limited by assumption of particle sphericity. The latter can be relaxed only if additional signals, such as LSP, are measured.
authors.affiliation: Voevodsky Institute of Chemical Kinetics and Combustion SB RAS, Institutskaya 3, 630090 Novosibirsk, Russia.
authors.forename: Anastasiya I
authors.initials: AI
authors.lastname: Konokhova
authors.affiliation: Voevodsky Institute of Chemical Kinetics and Combustion SB RAS, Institutskaya 3, 630090 Novosibirsk, RussiabNovosibirsk State University, Pirogova 2, 630090 Novosibirsk, Russia.
authors.forename: Darya N
authors.initials: DN
authors.lastname: Chernova
authors.affiliation: Voevodsky Institute of Chemical Kinetics and Combustion SB RAS, Institutskaya 3, 630090 Novosibirsk, RussiacNovosibirsk State Medical University, Krasny Prospect 52, 630091 Novosibirsk, Russia.
authors.forename: Dmitry I
authors.initials: DI
authors.lastname: Strokotov
authors.affiliation: State Research Institute of Circulation Pathology, Rechkunovskaya 15, 630055 Novosibirsk, Russia.
authors.forename: Andrei A
authors.initials: AA
authors.lastname: Karpenko
authors.affiliation: Voevodsky Institute of Chemical Kinetics and Combustion SB RAS, Institutskaya 3, 630090 Novosibirsk, RussiabNovosibirsk State University, Pirogova 2, 630090 Novosibirsk, Russia.
authors.forename: Andrei V
authors.initials: AV
authors.lastname: Chernyshev
authors.affiliation: Voevodsky Institute of Chemical Kinetics and Combustion SB RAS, Institutskaya 3, 630090 Novosibirsk, RussiabNovosibirsk State University, Pirogova 2, 630090 Novosibirsk, RussiacNovosibirsk State Medical University, Krasny Prospect 52, 630091 Novosibirsk, Russia.
authors.forename: Valeri P
authors.initials: VP
authors.lastname: Maltsev
authors.affiliation: Voevodsky Institute of Chemical Kinetics and Combustion SB RAS, Institutskaya 3, 630090 Novosibirsk, RussiabNovosibirsk State University, Pirogova 2, 630090 Novosibirsk, Russia.
authors.forename: Maxim A
authors.initials: MA
authors.lastname: Yurkin
mesh_terms: D055252:Cell-Derived Microparticles / Q000737:chemistry* / Q000502:physiology; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D006801:Humans; D008027:Light; D057230:Limit of Detection; D053657:Platelet-Rich Plasma / Q000166:cytology*; D012015:Reference Standards; D015203:Reproducibility of Results; D012542:Scattering, Radiation