A 10 μL drop of EVs in PBS was applied to nitrocellulose carbon-coated PELCO® Cu grids (Ted Pella Inc., Redding, CA, USA) and incubated for 1 min. Liquid was removed by touching the paper edge. A 10 μL drop of 2% uranyl acetate was immediately applied to the grid, followed by 15 s incubation and drop removal by touching the paper edge. Samples were examined at 80 kV with a JEM-1011 transmission electron microscope (JEOL, Akishima, Japan) equipped with an Orius™ SC1000 W camera (Gatan Inc., Pleasanton, CA, USA).
Orius sc1000 w camera
Manufactured by Ametek
Sourced in Japan
The Orius™ SC1000 W is a high-performance digital camera designed for scientific and industrial applications. It features a 16-megapixel CMOS sensor and provides high-quality imaging capabilities. The camera is capable of capturing detailed images and data for a variety of laboratory and research purposes.
Automatically generated - may contain errors
3 protocols using orius sc1000 w camera
1
Transmission Electron Microscopy Analysis of Extracellular Vesicles
2
Transmission Electron Microscopy of Biological Samples
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Ultrathin sections were examined by TEM. The material was fixed with 2% v/v glutaraldehyde in 0.1 mol l−1 cacodylate buffer (pH = 7.2). With Mycoplasma cultures, fixative was added (0.1 ml per 2 ml suspension) and centrifuged (Eppendorf AG, Hamburg, Germany) at 12 000g for 30 min, and 2 ml of fixative was added to the sediment obtained. When ejaculates were fixed, they were diluted 20-fold with isotonic NaCl solution, 0.1 ml of fixative per 2 ml of semen was added and centrifuged at 1500g (Elmi-tech) for 10 min, and 2 ml of fixative was added to the precipitate. All obtained samples were fixed with osmium tetroxide. After dehydration, they were embedded in epoxy according to standard techniques. Ultrathin sections were done on a Reichert Jung ultramicrotome, Ultracut E (Vienna, Austria), mounted on copper grids covered with a Formvar film, and contrasted with 1% aqueous uranyl acetate solution and lead citrate solution. The preparations were examined at 80 kV using a JEM-1011 transmission electron microscope (JEOL, Akishima, Japan) equipped with an Orius SC1000 W camera (Gatan Inc., Pleasanton, CA, USA).
3
Transmission Electron Microscopy of Extracellular Vesicles
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The carbon-coated TEM grids (Ted Pella, Redding, CA, USA) were treated using a glow discharge device Emitech K100X (Quorum Technologies Ltd., Lewes, UK) at 25 mA current for 45 s. This treatment turned the carbon surface hydrophilic and increased the EVs’ adsorption. The suspension of EVs was deposited onto the grid for 2–3 min, and the liquid was blotted using filter paper (OdiChem, Moscow, Russia). The grids were quickly transferred onto the drops of 1% uranyl acetate, stained for 1–2 min, blotted and dried. Images were obtained using a JEM-1011 (Jeol, Tokyo,Japan) transmission electron microscope equipped with Orius SC1000W camera (Gatan Inc., Pleasanton, CA, USA). The magnification was in the range from 30,000× to 250,000×; the acceleration voltage was 80 kV. The sizes of the EVs were measured using ScanEV [32 (link)].
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