Imaging of Isolated Extracellular Vesicles Using Fluorescence Microscopy

  • Dmitry Ter-Ovanesyan
  • Emma J. K. Kowal
  • Aviv Regev
  • George M. Church
  • Emanuele CocucciEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1660)


High-resolution fluorescence microscopy approaches enable the study of single objects or biological complexes. Single object studies have the general advantage of uncovering heterogeneity that may be hidden during the ensemble averaging which is common in any bulk conventional biochemical analysis. The implementation of single object analysis in the study of extracellular vesicles (EVs) may therefore be used to characterize specific properties of vesicle subsets which would be otherwise undetectable. We present a protocol for staining isolated EVs with a fluorescent lipid dye and attaching them onto a glass slide in preparation for imaging with total internal reflection fluorescence microscopy (TIRF-M) or other high-resolution microscopy techniques.

Key words

Exosomes Extracellular vesicles EVs Imaging Microscopy TIRF 



This work was supported by US National Institutes of Health National Human Genome Research Institute Grant P50 HG005550, The Ohio State Cancer Center (P30-CA016058), and an intramural fund (IRP46050-502339) granted to EC.


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Dmitry Ter-Ovanesyan
    • 1
    • 2
    • 3
    • 4
  • Emma J. K. Kowal
    • 1
    • 3
  • Aviv Regev
    • 4
    • 5
  • George M. Church
    • 1
    • 3
  • Emanuele Cocucci
    • 6
    Email author
  1. 1.Department of GeneticsHarvard Medical SchoolBostonUSA
  2. 2.Department of Molecular and Cellular BiologyHarvard UniversityBostonUSA
  3. 3.Wyss Institute for Biologically Inspired EngineeringHarvard UniversityBostonUSA
  4. 4.Broad Institute of MIT and HarvardCambridgeUSA
  5. 5.Department of BiologyMITCambridgeUSA
  6. 6.Division of Pharmaceutics and Pharmaceutical ChemistryCollege of Pharmacy, The Ohio State UniversityColumbusUSA

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