Related Experiment Video
Updated: Nov 18, 2025
09:01
Uptake of Fluorescent Labeled Small Extracellular Vesicles In Vitro and in Spinal Cord
Published on: May 23, 2021
3.9K
Fluorescent labeling of extracellular vesicles
Mehdi Dehghani1, Thomas R Gaborski1
1Department of Microsystems Engineering, Rochester Institute of Technology, Rochester, NY, United States; Department of Biomedical Engineering, Rochester Institute of Technology, Rochester, NY, United States.
Methods in Enzymology
|February 10, 2021
Summary
Fluorescent labeling of extracellular vesicles (EVs) aids in studying their cellular interactions and biodistribution. This guide reviews labeling methods, ideal dyes, and purification techniques, emphasizing size preservation for accurate EV research.
Area of Science:
- Biotechnology
- Cell Biology
- Nanotechnology
Background:
- Fluorescent labeling is crucial for tracking extracellular vesicles (EVs) in biological systems.
- Understanding EV uptake, biodistribution, and cellular effects requires precise labeling methods.
- High-resolution flow cytometry enables single EV-level characterization.
Purpose of the Study:
- To highlight the significance of fluorescent labeling for extracellular vesicle (EV) research.
- To review available fluorescent dyes, labeling strategies, and purification techniques.
- To assess the impact of labeling on EV size, critical for biodistribution and uptake studies.
Main Methods:
- Discussion of various fluorescent dyes and labeling approaches for EVs.
- Evaluation of ideal dye characteristics for EV labeling.
- Analysis of post-labeling purification techniques.
- Investigation of the effect of lipophilic (PKH) and luminal (CFSE) dyes on EV size.
- Presentation of a protocol for luminal EV labeling and size characterization using Nanoparticle Tracking Analysis (NTA).
Main Results:
- Common lipophilic and luminal dyes can alter the size of extracellular vesicles.
- Nanoparticle Tracking Analysis (NTA) is effective for characterizing EV size changes post-labeling.
- Preserving EV size is essential for accurate biodistribution and cellular uptake studies.
Conclusions:
- Appropriate selection of fluorescent dyes and labeling methods is vital for maintaining EV integrity.
- Careful characterization of labeling effects on EV size is necessary for reliable experimental outcomes.
- Optimized fluorescent labeling protocols enhance the utility of EVs in biological research.
