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Related Experiment Video

Updated: Nov 2, 2025

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
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Rapid Characterization and Quantification of Extracellular Vesicles by Fluorescence-Based Microfluidic Diffusion

Carolina Paganini1, Britta Hettich1, Marie R G Kopp1

  • 1Department of Chemistry and Applied Biosciences, ETH Zürich, Vladimir-Prelog-Weg 1-5/10, Zürich, 8093, Switzerland.

Advanced Healthcare Materials
|June 10, 2021
PubMed
Summary

This study introduces a microfluidic device for rapid extracellular vesicle (EV) characterization. The novel technique efficiently measures EV size, concentration, and composition, accelerating diagnostic and therapeutic development.

Keywords:
characterizationextracellular vesiclesfluorescenceimmunoassaymicrofluidicspurityquantification

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Analytical Chemistry

Background:

  • Extracellular vesicles (EVs) show promise as diagnostic and therapeutic agents.
  • Current EV characterization methods are time-consuming and material-intensive.
  • Standardization and efficiency in EV analysis are crucial for clinical translation.

Purpose of the Study:

  • To develop a rapid, integrated microfluidic device for comprehensive EV characterization.
  • To enable simultaneous measurement of EV size, concentration, and composition.
  • To improve the efficiency and reduce sample handling in EV analysis.

Main Methods:

  • A microfluidic device combining diffusion sizing and multiwavelength fluorescence detection.
  • Nonspecific staining for lipids/proteins and specific staining for EV markers (e.g., tetraspanins).
  • Integrated separation and quantification of free and EV-bound fluorophores for immunoassay functionality.

Main Results:

  • Simultaneous determination of EV size, concentration, and composition in minutes.
  • Detection and quantification of EV subtypes, impurities, and purity.
  • Minimal sample volume (<5 µL) and handling required, with direct in-solution analysis.

Conclusions:

  • The microfluidic device offers a rapid and efficient complementary tool for EV characterization.
  • This technology can accelerate screening, improve sample reproducibility assessment, and aid bioprocess development.
  • The method enhances the utility of EVs in diagnostics and therapeutics by streamlining their analysis.