I will be labelling isolated exosomes/EVs with the fluorescent dye sulfo-cy5.5 NHS ester for injection into mice. I was just wondering what the best way would be to check that my EVs have been appropriately and sufficiently labelled before injecting them into mice? For instance, could you place the labelled EVs into a multi-well plate and use a plate reader to check fluorescence? Or could you simply look at them under a microscope? Thank you!
Exosomes are too small to be seen by standard fluorescence microscopy. You would have to use ultraresolution microscopy techniques to resolve them, although you would probably be able to at least detect them as points by laser scanning confocal microscopy.
You could measure their fluorescence intensity using a fluorescence plate reader or a spectrofluorometer. To take fluorescence excitation and emission spectra, you could use a spectrofluorometer or a monochromator-based fluorescence plate reader. You could also run them on SDS-PAGE and detect the fluorescence of labeled proteins as bands using a fluorescence gel imager.
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