The established protocol of Bachurski et al. [50 (link)] was applied. Briefly, 5 µL of each sample was loaded onto formvar-coated copper grids (Science Services, Munich, Germany). After 20 min incubation, the grid-bound EVs were fixed with 2% paraformaldehyde for 5 min. Samples were washed with PBS and fixed again with 1% glutaraldehyde for 5 min, washed with Milli-Q water, and incubated with contrast dye (1.5% uranyl acetate) for 4 min. Images were acquired using a Gatan OneView 4 K camera (Gatan, Pleasanton, CA, USA) mounted on a Jem-2100Plus microscope (JEOL) operating at 200 kV.
Tzaridis T., Bachurski D., Liu S., Surmann K., Babatz F., Gesell Salazar M., Völker U., Hallek M., Herrlinger U., Vorberg I., Coch C., Reiners K.S, & Hartmann G. (2021). Extracellular Vesicle Separation Techniques Impact Results from Human Blood Samples: Considerations for Diagnostic Applications. International Journal of Molecular Sciences, 22(17), 9211.
Corresponding Organization : University Hospital Bonn
Other organizations :
Discovery Institute, Sanford Burnham Prebys Medical Discovery Institute, University Hospital Cologne, Cologne Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases, University of Cologne, German Center for Neurodegenerative Diseases, Universitätsmedizin Greifswald
Morphology and size of extracellular vesicles (EVs) as observed by transmission electron microscopy (TEM)
control variables
Formvar-coated copper grids used for sample application
Incubation time (20 min)
Fixation with 2% paraformaldehyde (5 min)
Fixation with 1% glutaraldehyde (5 min)
Contrast dye (1.5% uranyl acetate, 4 min)
Imaging using Gatan OneView 4 K camera on Jem-2100Plus microscope operating at 200 kV
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
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