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Imaging of Isolated Extracellular Vesicles Using Fluorescence Microscopy

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Extracellular Vesicles

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1660))

Abstract

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.

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Acknowledgment

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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Author notes

  1. Dmitry Ter-Ovanesyan and Emma J.K. Kowal contributed equally to this work.

Authors and Affiliations

  1. Department of Genetics, Harvard Medical School, Boston, MA, USA

    Dmitry Ter-Ovanesyan, Emma J. K. Kowal & George M. Church

  2. Department of Molecular and Cellular Biology, Harvard University, Boston, MA, USA

    Dmitry Ter-Ovanesyan

  3. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA

    Dmitry Ter-Ovanesyan, Emma J. K. Kowal & George M. Church

  4. Broad Institute of MIT and Harvard, Cambridge, MA, USA

    Dmitry Ter-Ovanesyan & Aviv Regev

  5. Department of Biology, MIT, Cambridge, MA, USA

    Aviv Regev

  6. Division of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy, The Ohio State University, Columbus, OH, USA

    Emanuele Cocucci

Authors
  1. Dmitry Ter-Ovanesyan
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  2. Emma J. K. Kowal
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  3. Aviv Regev
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  4. George M. Church
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  5. Emanuele Cocucci
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Corresponding author

Correspondence to Emanuele Cocucci .

Editor information

Editors and Affiliations

  1. CloudHealth Genomics, Ltd., Shanghai, China

    Winston Patrick Kuo

  2. Predicine, Inc., Hayward, California, USA

    Shidong Jia

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Ter-Ovanesyan, D., Kowal, E.J.K., Regev, A., Church, G.M., Cocucci, E. (2017). Imaging of Isolated Extracellular Vesicles Using Fluorescence Microscopy. In: Kuo, W., Jia, S. (eds) Extracellular Vesicles. Methods in Molecular Biology, vol 1660. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7253-1_19

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  • DOI: https://doi.org/10.1007/978-1-4939-7253-1_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7251-7

  • Online ISBN: 978-1-4939-7253-1

  • eBook Packages: Springer Protocols

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