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Related Experiment Video

Updated: Oct 31, 2025

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
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Multifluorescence Single Extracellular Vesicle Analysis by Time-Sequential Illumination and Tracking.

Siwoo Cho1,2, Johan Yi1, Yongmin Kwon1

  • 1Department of Mechanical Engineering, Pohang University of Science and Technology, 77 Cheongam-Ro. Nam-Gu, Pohang, Gyeong-buk 37673, Republic of Korea.

ACS Nano
|June 28, 2021
PubMed
Summary

This study introduces a new fluorescence nanoparticle tracking analysis (NTA) system to measure the size and membrane proteins of individual extracellular vesicles (EVs). This advanced NTA method aids in understanding individual EVs and their protein expression.

Keywords:
extracellular vesiclemultifluorescencenanoparticlenanoparticle tracking analysistime-sequential illumination

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Cell Biology

Background:

  • Extracellular vesicles (EVs) are crucial for intercellular communication.
  • Characterizing individual EVs, including their size and surface proteins, is vital for understanding their function.
  • Existing methods often lack the ability to analyze both size and protein expression simultaneously at the single-vesicle level.

Purpose of the Study:

  • To develop and validate a novel fluorescence-based nanoparticle tracking analysis (NTA) system.
  • To enable simultaneous characterization of extracellular vesicle (EV) size and membrane protein expression.
  • To advance the understanding of individual EV heterogeneity and function.

Main Methods:

  • A fluorescence-based nanoparticle tracking analysis (NTA) system utilizing a multi-wavelength laser sheet.
  • Sequential illumination with different laser wavelengths to capture scattering and fluorescence images.
  • Tracking of individual EVs via scattering images and detection of fluorescence-labeled membrane proteins.

Main Results:

  • The developed NTA system successfully characterized the size of individual extracellular vesicles (EVs).
  • Simultaneous detection of membrane protein expression (tetraspanins CD9, CD63, CD81) on individual EVs was achieved.
  • Analysis of HEK293 EVs demonstrated good correlation with established methods like total internal reflection fluorescence microscopy.

Conclusions:

  • The fluorescence-based NTA system provides a powerful tool for detailed characterization of individual EVs.
  • This method enhances the ability to study EV heterogeneity and protein expression profiles.
  • The system holds significant potential for advancing research in extracellular vesicle biology and diagnostics.