Copper/palladium grids (Agar Scientific) were covered with a 4 nm carbon-layer, performed in-house. Glow-discharge of grids was performed using a BAL-TEC sputter coater CD005 for 120 seconds. CeHUWE1 was diluted to 0.02 mg/ml in 10 mM HEPES, 100 mM NaCl pH 7.5 and applied to the grid for 60 seconds prior to staining with 2% uranyl acetate. A 200kV FEI Tecnai G2 20 (T20) transmission electron microscope was used to image CeHUWE1 grids at a 50,000x magnification (pixel size 2.211 Å). The micrographs obtained were processed using Relion 3.150 . Initial number of particles following auto-picking was ~65,000, reduced to ~14,000 at the final 2D classification, and ~9,000 for the final 3D model.
Tecnai g2 20 t20
Manufactured by Thermo Fisher Scientific
The Tecnai G2 20 (T20) is a transmission electron microscope (TEM) designed for high-resolution imaging and analysis of a wide range of samples. It features a 200 kV electron beam and advanced optics for delivering high-quality, high-contrast images. The T20 is capable of resolving fine structural details at the nanometer scale.
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Lab products found in correlation
2 protocols using tecnai g2 20 t20
1
Cryo-EM structure determination of CeHUWE1
2
Cryo-EM structure determination of CeHUWE1
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Copper/palladium grids (Agar Scientific) were covered with a 4 nm carbon-layer, performed in-house. Glow-discharge of grids was performed using a BAL-TEC sputter coater CD005 for 120 seconds. CeHUWE1 was diluted to 0.02 mg/ml in 10 mM HEPES, 100 mM NaCl pH 7.5 and applied to the grid for 60 seconds prior to staining with 2% uranyl acetate. A 200kV FEI Tecnai G2 20 (T20) transmission electron microscope was used to image CeHUWE1 grids at a 50,000x magnification (pixel size 2.211 Å). The micrographs obtained were processed using Relion 3.150 . Initial number of particles following auto-picking was ~65,000, reduced to ~14,000 at the final 2D classification, and ~9,000 for the final 3D model.
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