Ccd 2 k 2 k veleta model

sEV morphology was analysed in two sEV samples obtained from plasma and CSF. The morphology was evaluated by transmission electron microscopy (TEM) using the methodology previously described [75 (link),76 (link)]. A volume of 5 µL of resuspended sEVs were fixed in 2.5% glutaraldehyde and then washed with deionised water. Samples were contrasted with 2% uranyl acetate, embedded in 0.13% methyl cellulose and 0.4% uranyl acetate, and then visualised using a Tecnai T20 microscope (FEI Company, Hillsboro, OR, USA), with a filament of LaB6. The voltage used during the visualisation was 200 KV, and acquiring images was performed with a CCD 2 K × 2 K Veleta model (Olympus, Tokyo, Japan).

Exosome morphology was evaluated by TEM using the methodology previously described by Lötvall et al.⁵⁰ . First, total protein in each sample was evaluated by BCA (B9643, C2284, Sigma). A drop of 10 μg of exosomal protein was placed on a Parafilm layer. Exosomes were fixed in 2.5% glutaraldehyde and then washed with deionized water. Samples were contrasted with 2% uranyl acetate, embedded in 0.13% methyl cellulose and 0.4% uranyl acetate and then visualized using a Tecnai T20 microscope (FEI Company), with a filament of LaB₆. The voltage used during the visualization was 200 KV, and acquiring images was performed with a CCD 2 K × 2 K Veleta model (Olympus).