Fluorescent labeling of extracellular vesicles
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Updated: Nov 18, 2025
Uptake of Fluorescent Labeled Small Extracellular Vesicles In Vitro and in Spinal Cord
Published on: May 23, 2021
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.
Abstract:
Fluorescent labeling of extracellular vesicles (EVs) enables studying their uptake and influence on individual cells, biodistribution as well as facilitates their characterization using high-resolution flow cytometry at a single EV level. Here we describe the importance of fluorescent labeling, the available fluorescent dyes and labeling approaches, the characteristics of an ideal dye, and the available techniques for post-labeling purification. We discuss the importance of preserving the size of EVs for uptake, biodistribution, and characterization studies and focus on the effect of common lipophilic PKH and luminal CFSE dyes on the size of EVs. Lastly, we present an example protocol for luminal labeling of EVs and characterization of the effect of labeling on the size of EVs using nanoparticles tracking analysis (NTA).
