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Updated: May 11, 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
Nanoscale Fluorescent Characterizations and Origin Tracing of Extracellular Vesicles through Standard Preparation
Herath D W Herath1, Haoran Jing1, Mingyang Li2
1Department of Chemistry, College of Liberal Arts and Sciences, University of Illinois Chicago, Chicago, Illinois 60607, United States.
ACS Nano
|May 9, 2026
Summary
This study uses direct stochastic optical reconstruction microscopy (dSTORM) to analyze extracellular vesicles (EVs) at the nanoscale. Standardized EV isolation and dSTORM imaging reveal sub-EV heterogeneity and biomarker distribution for improved diagnostics.
Area of Science:
- Biotechnology
- Cell Biology
- Nanotechnology
Background:
- Extracellular vesicles (EVs) are crucial for intercellular communication and serve as biomarkers and therapeutic agents.
- EV heterogeneity necessitates sub-EV resolution analysis for precise profiling and understanding biogenesis.
- Current limitations in EV purification, immobilization, and labeling hinder reproducibility and interpretation.
Purpose of the Study:
- To develop and validate a robust workflow for nanoscale characterization of EVs using direct stochastic optical reconstruction microscopy (dSTORM).
- To investigate the nanoscale spatial organization of CD81 on EVs from different cellular sources.
- To differentiate between EV subtypes (ectosomes and exosomes) using high-resolution imaging.
Main Methods:
- Isolation of EVs from cultured cells using polymer-based precipitation and size-exclusion chromatography.
- Immobilization of purified EVs on poly-l-lysine-coated surfaces for immunofluorescence staining.
- Application of dSTORM super-resolution microscopy for nanoscale imaging and quantitative analysis of EV surface markers.
Main Results:
- dSTORM imaging revealed broad EV size distributions and variations in nanoscale CD81 organization.
- A subpopulation of EVs with asymmetrical CD81 distribution, below the optical diffraction limit, was identified and quantified.
- Successful distinction between ectosomes and exosomes was achieved using pan-membrane-protein labeling.
Conclusions:
- Coupling standard EV isolation methods with dSTORM provides a powerful strategy for high-resolution sub-EV analysis.
- This approach enables precise characterization of EV heterogeneity and facilitates biomarker discovery.
- The developed workflow enhances reproducibility and biological interpretation of EV studies.
