Can anyone suggest a protocol (with reference) on how to prepare the exosomes for CONFOCAL STAINING? Details- exosmes will be isolated from culture media at 100,000g and will be resuspended in PBS. Now--- how do I fix it and ready it for confocal?
Do we need to fix at all? The labels for exosome I have in mind are antibodies OR organic dyes.
Thanks!
Hello Farah, are you using ultra-resolution microscopy for your EVs? Like STORM microscope? regular confocal may be challenging to do due to EV's size. The key would be to immobilize the EVs on the substrate. You can use something like polyLysine coating as it will provide a positive charge on the substrate and help fix particles in place. It may be easier to label particles in solution, and then just deposit on your slide. A good alternative or complementary method may be multicolor fluorescent nanoparticle tracking analysis for quantitative results or immunogold TEM.
Good luck!
Farah - How could you image Exosomes by confocal ?? Do you really think you can?
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