Carbon coated 200 cu mesh grid

Manufactured by Electron Microscopy Sciences

The Carbon-coated 200 Cu mesh grid is a laboratory equipment designed for use with electron microscopes. It provides a substrate for holding and supporting samples during imaging and analysis. The grid features a copper (Cu) mesh structure coated with a thin layer of carbon, which helps to improve the conductivity and stability of the sample during electron beam exposure.

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Lab products found in correlation

Fei t20 electron microscope, by Thermo Fisher Scientific (1 mentions) Fab preparation kit, by Thermo Fisher Scientific (1 mentions) Eagle 2kx2k ccd camera, by Thermo Fisher Scientific (1 mentions) Pelco easiglow, by Ted Pella (1 mentions) Whatman no 1 qualitative filter paper, by Cytiva (1 mentions) Pelco easiglow glow discharge cleaning system, by Ted Pella (1 mentions)

2 protocols using carbon coated 200 cu mesh grid

Structural analysis of GPC2-CT3 Fab complex

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The CT3 antigen-binding fragment (Fab) was prepared using a Fab preparation kit (Thermo Scientific). GPC2 protein was mixed with CT3 Fab at 1:1 molar ratio in PBS. A 3 μl aliquot containing ∼0.01 mg/ml of the sample was applied for 20 s onto a carbon-coated 200 Cu mesh grid (Electron Microscopy Sciences, Protochips, Inc.) that had been glow discharged at 30 mA for 30 s (Pelco easiGlow, Ted Pella, Inc.), then negatively stained with 0.7% (w/v) uranyl formate for 40 s. Data was collected using a Tecnai FEI T20 electron microscope operating at 200 kV, with an electron dose of ∼40 e-/Å2 and a magnification of 100,000 x that resulted in a pixel size of 2.19 Å at the specimen plane. Images were acquired with an Eagle 2kx2k CCD camera (FEI) using a nominal defocus of 1100 nm and the SerialEM software.⁴¹ (link)

Li N., Torres M.B., Spetz M.R., Wang R., Peng L., Tian M., Dower C.M., Nguyen R., Sun M., Tai C.H., de Val N., Cachau R., Wu X., Hewitt S.M., Kaplan R.N., Khan J., St Croix B., Thiele C.J, & Ho M. (2021). CAR T cells targeting tumor-associated exons of glypican 2 regress neuroblastoma in mice. Cell Reports Medicine, 2(6), 100297.

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Negative Staining EM Grid Preparation

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Negatively stained EM grids were prepared from the purified samples at a concentration of 0.08 mg/μl. 3 μl of diluted sample was applied to carbon-coated 200 Cu mesh grid (Electron Microscopy Sciences, Inc.) with pretreated glow discharge at 15mA for 15 seconds by a Pelco easiGlow Glow discharge cleaning system (Ted Pella, Inc.) and left as such for 20 seconds. 3 μl of 0.7% (w/v) uranyl formate was added for 5 seconds followed by blotting using Whatman No. 1 qualitative filter paper. This staining process was repeated another 4 times. Samples were imaged on Talso 120C TEM (ThermoFisher Scientific) that operated in 120 kV integrated with 4k x 4k CCD. Micrographs were collected using SerialEM software (Mastronarde, 2005 (link)) with magnification of 36,000 x that resulted in a pixel size of 2.94 Å at the specimen plane.

Arango D., Sturgill D., Yang R., Kanai T., Bauer P., Roy J., Wang Z., Hosogane M., Schiffers S, & Oberdoerffer S. (2022). Direct epitranscriptomic regulation of mammalian translation initiation through N4-acetylcytidine. Molecular cell, 82(15), 2797-2814.e11.

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