400 mesh copper grids with a supporting carbon lacey film coated with an ultra-thin carbon support film <3 nm thick (Agar Scientific, UK) were employed. Grids were glow-discharged for 30 s (easiGlow, Ted Pella) prior to applying 3 μl of purified ribosomes, and vitrification was performed by plunge-freezing in liquid ethane cooled by liquid nitrogen using a Leica EM GP device (Leica Microsystems). Samples were diluted using the buffer exchange buffer (50 mM Tris–HCl pH 8, 150 mM NaCl, 10 mM MgCl2) as required. Cryo-EM data was collected on a FEI Titan Krios (Astbury Biostructure Laboratory, University of Leeds) EM at 300 kV, using a total electron dose of 80 e–/Å2 and a magnification of 75 000× at –2 to –4 μm defocus. Movies were recorded using the EPU automated acquisition software on a FEI Falcon III direct electron detector, in linear mode, with a final pixel size of 1.065 Å/pixel (Supplementary Table S2 ).
Ultra thin carbon support film
Manufactured by Agar Scientific
Sourced in United Kingdom
Ultra-thin carbon support film is a laboratory equipment used to provide a thin, electron-transparent substrate for the mounting and analysis of samples in electron microscopy. It serves as a support structure for the samples, facilitating their examination under the microscope.
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Lab products found in correlation
2 protocols using ultra thin carbon support film
1
Cryo-EM Sample Preparation Protocol
2
Cryo-EM Imaging of Purified Ribosomes
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400 mesh copper grids with a supporting carbon lacey film coated with an ultra-thin carbon support film < 3 nm thick (Agar Scientific, UK) were employed. Grids were glowdischarged for 30 seconds (easiGlow, Ted Pella) prior to applying 3 µL of purified ribosomes, and vitrification was performed by plunge-freezing in liquid ethane cooled by liquid nitrogen using a Leica EM GP device (Leica Microsystems). Samples were diluted using the buffer exchange buffer (50 mM Tris-HCl pH 8, 150 mM NaCl, 10 mM MgCl2) as required. Cryo-EM data was collected on a FEI Titan Krios (Astbury Biostructure Laboratory, University of Leeds) EM at 300 kV, using a total electron dose of 80 e -/Å 2 and a magnification of 75,000 × at -2 to -4 μm defocus. Movies were recorded using the EPU automated acquisition software on a FEI Falcon III direct electron detector, in linear mode, with a final pixel size of 1.065 Å/pixel (Sup Table 2).
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