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Related Experiment Video

Updated: Oct 2, 2025

Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis
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Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis.

Chi-Ju Kim1, Morgan D Kuczler2, Liang Dong3

  • 1The Brady Urological Institute, Johns Hopkins University School of Medicine; Department of Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST); ckim143@jh.edu.

Journal of Visualized Experiments : Jove
|February 28, 2022
PubMed
Summary

This study introduces a practical assay for quantifying extracellular vesicle (EV) uptake in cells. The method uses 3D confocal microscopy to accurately measure internalized EVs, advancing cell communication research.

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Area of Science:

  • Cell Biology
  • Biotechnology
  • Microscopy

Background:

  • Extracellular vesicle (EV) uptake is crucial for intercellular communication in fields like cancer biology, neuroscience, and drug delivery.
  • Existing EV uptake assays often lack detailed, practical experimental methodologies, hindering accurate quantification.
  • Distinguishing internalized EVs from superficial ones is a significant challenge in current assays.

Purpose of the Study:

  • To develop a practical and robust assay for visualizing and quantifying cellular extracellular vesicle (EV) uptake.
  • To address the limitations of existing methods in accurately distinguishing internalized EVs from those on the cell surface.
  • To provide a detailed experimental protocol for efficient EV uptake analysis at the cellular level.

Main Methods:

  • Preparation of fluorescently labeled extracellular vesicles (EVs) using a nano-filtration-based microfluidic device.
  • Visualization of labeled EVs within cells using three-dimensional (3D) fluorescence confocal microscopy.
  • Quantitative analysis of EV uptake through advanced image-processing software.

Main Results:

  • A robust methodology for analyzing EV uptake at the cellular level was established.
  • The assay enables accurate quantification of internalized EVs by distinguishing them from superficial EVs.
  • The developed protocol offers a practical approach for efficient EV uptake analysis.

Conclusions:

  • The proposed 3D confocal microscopy-based assay provides a practical and efficient method for quantifying EV uptake.
  • This assay overcomes challenges in distinguishing internalized from superficial EVs, improving accuracy in cellular communication studies.
  • The detailed protocol supports advancements in research areas relying on understanding EV-mediated intercellular communication.