Tecnai g2 transmission electron

Freshly isolated mouse hippocampus samples were fixed overnight in a solution of 2.5% (v/v) glutaraldehyde plus 2% (v/v) paraformaldehyde in 100 mM sodium cacodylate (pH 7.2) at 4 °C. After the samples had been washed, postfixation was performed in a 1% OsO₄ solution in 100 mM sodium cacodylate (pH 7.2) at 4 °C. Sections were contrasted with a saturated uranyl acetate solution in 50% ethanol for 15 min, followed by incubation in a 0.5% (w/v) lead citrate solution in distilled water for 7 min. Finally, the samples were analyzed with a Tecnai G2 transmission electron microscope (FEI Company, Hillsboro, OR, USA) operating at 100 kV. Images were acquired with a Veleta camera (Olympus Soft Imaging Solutions, Italy).

EVs were observed by a Tecnai G2 transmission electron microscope (FEI, Hillsboro, USA). Samples were fixed with 1% glutaraldehyde, applied onto a carbon-coated copper grid, and stained with 1% phosphotungstic acid. NTA was performed using a ZetaView system (Particle Metrix, Meerbusch, Germany) to track the Brownian motion of EVs suspended in PBS, and size distribution data was generated by applying the Stokes-Einstein equation.