Research Article
Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis

https://doi.org/10.1016/j.nano.2011.04.003Get rights and content
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Abstract

Cellular microvesicles and nanovesicles (exosomes) are involved in many disease processes and have major potential as biomarkers. However, developments in this area are constrained by limitations in the technology available for their measurement. Here we report on the use of fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles. In this system vesicles are visualized by light scattering using a light microscope. A video is taken, and the NTA software tracks the brownian motion of individual vesicles and calculates their size and total concentration. Using human placental vesicles and plasma, we have demonstrated that NTA can measure cellular vesicles as small as ∼50 nm and is far more sensitive than conventional flow cytometry (lower limit ∼300 nm). By combining NTA with fluorescence measurement we have demonstrated that vesicles can be labeled with specific antibody-conjugated quantum dots, allowing their phenotype to be determined.

From the Clinical Editor

The authors of this study utilized fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles, demonstrating that NTA is far more sensitive than conventional flow cytometry.

Graphical Abstract

Screen shot from video of light scatter of placental vesicles overlaid with a graph of vesicle size and concentration as determined by nanoparticle tracking analysis.

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Key words

Microvesicles
Microparticles
Exosomes
Nanoparticle Tracking Analysis
Flow Cytometry

Cited by (0)

This work was supported by a Wellcome Trust Technology Development Grant (ref GR087730), Wellcome Trust Programme Grant (ref GR079862MA), and by the Oxford Partnership Comprehensive Biomedical Research Centre with funding from the Department of Health's NIHR Biomedical Research Centres funding scheme. The views expressed in this publication are those of the authors and not necessarily those of the Department of Health.

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These authors contributed equally to this work.