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Methods Mol Biol. 2017;1660:233-241. doi: 10.1007/978-1-4939-7253-1_19.

Imaging of Isolated Extracellular Vesicles Using Fluorescence Microscopy.

Author information

1
Department of Genetics, Harvard Medical School, Boston, MA, USA.
2
Department of Molecular and Cellular Biology, Harvard University, Boston, MA, USA.
3
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.
4
Broad Institute of MIT and Harvard, Cambridge, MA, USA.
5
Department of Biology, MIT, Cambridge, MA, USA.
6
Division of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy, The Ohio State University, Columbus, OH, USA. cocucci.1@osu.edu.

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.

KEYWORDS:

EVs; Exosomes; Extracellular vesicles; Imaging; Microscopy; TIRF

PMID:
28828661
DOI:
10.1007/978-1-4939-7253-1_19
[Indexed for MEDLINE]

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