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Updated: Feb 24, 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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Detection of Organelle-Specific Dyes Labeled Extracellular Vesicles with Colocalization-Fluorescence Nanoparticle
Getnet Midekessa1,2, Kasun Godakumara2, Mohammad Mehedi Hasan3
1Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu 50411, Estonia.
ACS Omega
|February 23, 2026
Summary
Researchers developed a new method using fluorescence and colocalization Nanoparticle Tracking Analysis (NTA) to trace extracellular vesicle (EV) origins. This technique enhances the study of single EVs and their role in cell communication.
Area of Science:
- Extracellular vesicle (EV) research
- Cellular biology
- Biotechnology
Background:
- Cell types release diverse extracellular vesicles (EVs) crucial for intercellular communication.
- Existing methods for analyzing heterogeneous EVs are limited in identifying individual vesicle origins and characteristics.
- Understanding EV biogenesis and intracellular origins is key to deciphering their biological roles.
Purpose of the Study:
- To develop and apply a novel fluorescence and colocalization Nanoparticle Tracking Analysis (NTA) method.
- To identify the subcellular origins of EVs from human choriocarcinoma cells (JAr) and bovine follicular fluids (BFF).
- To determine physical characteristics and colocalization ratios of endoplasmic reticulum (ER) and mitochondria (Mito)-positive EVs.
Main Methods:
- Utilized fluorescence and colocalization Nanoparticle Tracking Analysis (F-NTA).
- Employed size exclusion chromatography (SEC) for EV purification.
- Applied specific fluorescent dyes for labeling ER, mitochondria, and cell membranes (CMDR).
Main Results:
- Achieved high labeling efficiency for ER-positive JAr EVs (67.11%) and BFF EVs (96.27%) via SEC.
- Observed higher Mito-positive JAr EVs (25.74%) compared to BFF EVs (14.21%).
- Colocalization analysis revealed potential interactions between organelles and EV biogenesis pathways.
Conclusions:
- The integrated colocalization technology in F-NTA significantly advances single EV analysis.
- This method provides deeper insights into EV biology and their intracellular origins.
- The findings pave the way for more precise characterization of EVs in various biological contexts.
