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A liposome-based size calibration method for measuring microvesicles by flow cytometry

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  • Jens Bæk Simonsen

BACKGROUND: Over the last years the need for a gold standard to determine the sizes of extracellular vesicles including microvesicles by flow cytometry has been emphasized.

METHODS: This work suggests to use artificial vesicles as calibrators to ascertain the size of microvesicles from the side scattering (SSC) measured by flow cytometry. We prepared fluorescently labeled liposomes with different maximum sizes defined by the pore size (200, 400, 800, and 1000 nm) of the membrane used for the extrusion. The fluorescence strengths from the largest liposomes pertaining to each pore size enabled us to verify the correlation between the SSC from a liposome and the corresponding size.

CONCLUSIONS: We show that artificial vesicles are more accurate size calibrators compared to the commonly used polystyrene calibrator beads illustrated by the SSC from 110 nm polystyrene beads corresponds to the scattering from ~400 nm sized vesicle-like particles. We also show that this method of size assessment based on SSC has a low resolution that is roughly estimated to be between 60 to 200 nm dependent on the vesicle size. This article is protected by copyright. All rights reserved.

OriginalsprogEngelsk
TidsskriftJournal of Thrombosis and Haemostasis
Vol/bind14
Udgave nummer1
Sider (fra-til)186-190
Antal sider5
ISSN1538-7933
DOI
StatusUdgivet - 2016

ID: 147927585