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Hollow organosilica beads as reference particles for optical detection of extracellular vesicles
Author(s) -
Varga Z.,
Pol E.,
Pálmai M.,
GarciaDiez R.,
Gollwitzer C.,
Krumrey M.,
Fraikin J.L.,
Gasecka A.,
Hajji N.,
Leeuwen T. G.,
Nieuwland R.
Publication year - 2018
Publication title -
journal of thrombosis and haemostasis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.947
H-Index - 178
eISSN - 1538-7836
pISSN - 1538-7933
DOI - 10.1111/jth.14193
Subject(s) - materials science , polystyrene , flow cytometry , dispersity , light scattering , dynamic light scattering , cytometry , extracellular vesicles , nanotechnology , biomedical engineering , chromatography , chemistry , scattering , nanoparticle , polymer , optics , composite material , polymer chemistry , medicine , genetics , physics , microbiology and biotechnology , biology
Essentials Standardization of extracellular vesicle (EV) measurements by flow cytometry needs improvement. Hollow organosilica beads were prepared, characterized, and tested as reference particles. Light scattering properties of hollow beads resemble that of platelet‐derived EVs. Hollow beads are ideal reference particles to standardize scatter flow cytometry research on EVs.Summary Background The concentration of extracellular vesicles ( EV s) in body fluids is a promising biomarker for disease, and flow cytometry remains the clinically most applicable method to identify the cellular origin of single EV s in suspension. To compare concentration measurements of EV s between flow cytometers, solid polystyrene reference beads and EV s were distributed in the first ISTH ‐organized interlaboratory comparison studies. The beads were used to set size gates based on light scatter, and the concentration of EV s was measured within the size gates. However, polystyrene beads lead to false size determination of EV s, owing to the mismatch in refractive index between beads and EV s. Moreover, polystyrene beads gate different EV sizes on different flow cytometers. Objective To prepare, characterize and test hollow organosilica beads ( HOB s) as reference beads to set EV size gates in flow cytometry investigations. Methods HOB s were prepared with a hard template sol‐gel method, and extensively characterized for morphology, size, and colloidal stability. The applicability of HOB s as reference particles was investigated by flow cytometry with HOB s and platelet‐derived EV s. Results HOB s proved to be monodisperse with a homogeneous shell thickness. Two‐angle light‐scattering measurements by flow cytometry confirmed that HOB s have light‐scattering properties similar to those of platelet‐derived EV s. Conclusions Because the structure and light‐scattering properties HOB s resemble those of EV s, HOB s with a given size will gate EV s of the same size. Therefore, HOB s are ideal reference beads with which to standardize optical measurements of the EV concentration within a predefined size range.