Tecnai t12 g2 twin transmission electron microscope

EVs were isolated from pancreatic cancer cell line harboring mutp53 (PANC-1) utilizing ultracentrifugation followed by SEC. FEI Tecnai T12 G2 TWIN transmission electron microscope (Hillsboro, OR, USA) operating at 120 kV was used. The images were taken with Gatan 794 MultiScan CCD camera. In short, the samples were prepared in the following manner: 2.5 µL of the sample was applied on to 300 mesh copper grid and the excess liquid was blotted with filter paper after 1 min. The grid was dried in air for 1 min, followed by applying 5 µL of uranyl acetate 2% for negative staining in order to increase the sample contrast. Next, the grid was blotted once more in order to remove the excess uranyl acetate. Finally, the grid was dried in air before insertion in to the microscope.

Cultures (2 ml) of parent and ΔaglB stain cells were pelleted (2 min at 8000 × g in a microfuge) and the supernatant was removed. The pellet was carefully resuspended in 1 ml basal salt solution (corresponding to Hbt. salinarum CM without peptone). The resulting cell suspension (1 ml) was pelleted as before, the supernatant was removed, and the pellet was resuspended in 1 ml of BSS.
Aliquots (2.5 μl) were applied to 300 mesh copper grid and any excess liquid was blotted with filter paper after 1 min. The grid was dried in air for 1 min, when 5 μl of uranyl acetate (2%) was applied for negative staining to increase the sample contrast. Next, the grids were blotted once more to remove excess uranyl acetate. Finally, the grids were dried in air before insertion into the microscope. Electron micrographs were taken with a FEI Tecnai T12 G² TWIN transmission electron microscope operating at 120 kV.