A standarized protocol for evaluation of large extracellular vesicles using the attune™ NXT system

Extracellular vesicles (EVs) represent a promising biomarker in several medical areas. Flow cytometry (FC) is one of the most widely-used methods to characterize EVs, providing quantitative information and determination of EV subtypes. EV evaluation represents a challenge as no standardized methods...

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Bibliographic Details
Published in:Journal of immunological methods 2021-12, Vol.499, p.113170-113170, Article 113170
Main Authors: Lugo-Gavidia, Leslie Marisol, Burger, Dylan, Bosio, Erika, Robinson, Sandi, Nolde, Janis, Carnagarin, Revathy, Matthews, Vance B., Schlaich, Markus P.
Format: Article
Language:English
Subjects:
Attune Nxt
Extracellular vesicles
Flow cytometry
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Summary:Extracellular vesicles (EVs) represent a promising biomarker in several medical areas. Flow cytometry (FC) is one of the most widely-used methods to characterize EVs, providing quantitative information and determination of EV subtypes. EV evaluation represents a challenge as no standardized methods are available to facilitate assessment across different research centers. This is principally because their size falls below the detection limit of most standard flow cytometers and a thorough optimization process is required to ensure instrument-specific sensitivity. We provide an overview of a standardized method to evaluate large EVs using the Attune™ Nxt Acoustic Focusing Flow Cytometer system (Thermo Fisher Scientific). •Increased dynamic detection range and better discrimination on the SSC by the Attune™ NxT Small Particle Side-Scatter Filter.•Size approximation and internal control is achieved by the use of silica spheres of pre-specified diameters incorporating fluorescence.•The lowest throughput rate is critical to ensure sensitivity in extracellular vesicle characterization.
ISSN:0022-1759
1872-7905
DOI:10.1016/j.jim.2021.113170