Electron microscopy
Electron microscopy was carried out from 2µg of EVs that were fixed in
PBS-formaldehyde 2% (v/v) for 16 hours at 4°C and shaking. Particles
were adsorbed on copper-carbon grids (copper-formwar/carbon-coated grids
200 mesh, Fedelco) for 3 minutes, at room temperature (RT), and were
contrasted with uranyl acetate at 1% (v/v) for 30 seconds. Samples were
observed in a JEOL 1010 transmission electron microscope (JEOL), using a
working voltage of 80kV. The analysis was carried out at the Electron
Microscopy Center in the Universidad Complutense of Madrid (UCM), using
the Soft Imaging Viewer program.