Imaging of Isolated Extracellular Vesicles Using Fluorescence Microscopy
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 wordsExosomes 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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