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Updated: Jun 12, 2026
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Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
Published on: January 7, 2019
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Fluorescence Characterization of Extracellular Vesicles Using Single-Molecule Confocal Microscopy
Tianxiao Zhao1,2, Noelia Pelegrina-Hidalgo1,2, Daniel C Edwards1,2
1EaStCHEM School of Chemistry, The University of Edinburgh, Edinburgh, EH9 3FJ, UK.
Small Methods
|August 18, 2025
Summary
This study presents a new method for detecting and quantifying extracellular vesicles (EVs) using fluorescently labeled antibodies and advanced microscopy. The technique achieves high sensitivity, enabling EV biomarker discovery in biological samples.
Area of Science:
- Biotechnology
- Cell Biology
- Biomarker Discovery
Background:
- Extracellular vesicles (EVs) are crucial for intercellular communication and are emerging as significant biomarkers.
- Conventional methods struggle with the low abundance and heterogeneity of EVs, hindering accurate characterization.
- Specific detection of individual EVs is essential for reliable biomarker analysis.
Purpose of the Study:
- To develop a highly sensitive and specific method for detecting and quantifying individual extracellular vesicles (EVs).
- To enable the characterization of EVs directly in complex biological fluids like serum and plasma.
- To compare EV extraction yields from different purification methods.
Main Methods:
- Coupling EV-specific antibodies with two different fluorophores.
- Utilizing fast-flow microfluidics and single-molecule confocal microscopy for EV detection.
- Comparing ultracentrifugation and size exclusion chromatography for EV yield.
Main Results:
- Achieved sub-femtomolar detection limits (≈10^7 EVs mL^-1) for individual EVs.
- Demonstrated specific detection of EVs amidst other lipid vesicles.
- Quantified EVs in serum and plasma without prior purification.
- Compared EV yields from different isolation techniques.
Conclusions:
- The developed method offers a highly specific, sensitive, and user-friendly solution for EV characterization.
- This approach facilitates robust EV biomarker discovery and quantification in clinical samples.
- The technique overcomes limitations of conventional methods for analyzing heterogeneous EV populations.
