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Carbon coated 400 mesh copper grids

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Carbon-coated 400 mesh copper grids are a type of laboratory equipment used for sample preparation in electron microscopy. They provide a thin, conductive surface for mounting and supporting samples during imaging.

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Lab products found in correlation

Uranyl acetate, by ProSciTech (2 mentions) Uranyl acetate, by Merck Group (2 mentions) Glutaraldehyde, by Merck Group (2 mentions) Methylcellulose, by Merck Group (2 mentions) Valeta 4 mp ccd camera, by EMSIS (2 mentions) Jem 2100 transmission electron microscope, by JEOL (2 mentions) Uranyl acetate, by Agar Scientific (2 mentions) K350 glow discharge unit, by Quorum Technologies (2 mentions) Jem 2100 electron microscope, by JEOL (1 mentions)

4 protocols using carbon coated 400 mesh copper grids

1

TEM Sample Preparation for BMVs

Cited 1 time
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Sample preparation for TEM was performed as described previously (11 (link)). Briefly, BMVs were coated onto carbon-coated 400 mesh copper grids (ProSciTech), fixed in 1% (wt/vol) glutaraldehyde (Sigma) in PBS, stained with 2% (wt/vol) uranyl acetate (ProSciTech) (pH 7.0), and coated with 2% (wt/vol) methyl-cellulose (Sigma) in 0.4% (wt/vol) uranyl acetate (pH 4.0). Samples were viewed using a JEOL JEM-2100 transmission electron microscope (JEOL, Japan) operated at 200 kV fitted with a Valeta 4 MP CCD camera (Emsis, Germany).
Bitto N.J., Zavan L., Johnston E.L., Stinear T.P., Hill A.F, & Kaparakis-Liaskos M. (2021). Considerations for the Analysis of Bacterial Membrane Vesicles: Methods of Vesicle Production and Quantification Can Influence Biological and Experimental Outcomes. Microbiology Spectrum, 9(3), e01273-21.
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2

Negative Staining Protocol for EV Imaging

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EV samples (5 µl at 0.05 µg/ml protein) were deposited onto carbon-coated 400-mesh copper grids (ProSciTech) that had been glow discharged for 1 min in a K950X turbo evaporator coupled to a K350 glow discharge unit (Quorum Technologies Ltd), incubated for 1 min then washed with ultrapure water. Grids were then stained three times with 4 µl of 2% (v/v) uranyl acetate (Agar Scientific). Excess solution was blotted off and the grids were dried overnight. Images were captured using a JEM 2100 electron microscope (JEOL Ltd) operated at 200 kV.
Bleackley M.R., Samuel M., Garcia-Ceron D., McKenna J.A., Lowe R.G., Pathan M., Zhao K., Ang C.S., Mathivanan S, & Anderson M.A. (2020). Extracellular Vesicles From the Cotton Pathogen Fusarium oxysporum f. sp. vasinfectum Induce a Phytotoxic Response in Plants. Frontiers in Plant Science, 10, 1610.
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3

TEM Imaging of Outer Membrane Vesicles

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TEM sample preparation was performed as previously described (5 (link), 35 (link)). Briefly, OMVs were coated onto carbon‐coated 400 mesh copper grids (ProSciTech, Australia) for 10 min, fixed in 1% (w/v) glutaraldehyde (Sigma-Aldrich, USA) and negatively-stained with 2% (w/v) uranyl-acetate (ProSciTech, Australia). OMV samples were then coated with 2% (w/v) methyl-cellulose (Sigma-Aldrich, USA) in 0.4% (w/v) uranyl acetate. Samples were air dried and viewed using a JEM-2100 transmission electron microscope (JEOL, Japan) operated at 200 kV using a Valeta 4 MP CCD camera (Emsis, Germany).
Gilmore W.J., Johnston E.L., Bitto N.J., Zavan L., O'Brien-Simpson N., Hill A.F, & Kaparakis-Liaskos M. (2022). Bacteroides fragilis outer membrane vesicles preferentially activate innate immune receptors compared to their parent bacteria. Frontiers in Immunology, 13, 970725.
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4

Negative Staining of Extracellular Vesicles

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Carbon-coated 400-mesh copper grids (ProSciTech) were glow discharged for 1 min in a K950X turbo evaporator coupled to a K350 glow discharge unit (Quorum Technologies Ltd). EVs (5 µL) were deposited onto the grid and incubated for 1 min. Grids were then washed once with ultrapure water and negatively stained three times with 2% (v/v) uranyl acetate (Agar Scientific). Excess solution was blotted off and the grids were dried overnight. AJEM 2100 electron microscope (JEOL Ltd) operated at 200 kV was used for imaging.
Dawson C.S., Garcia-Ceron D., Rajapaksha H., Faou P., Bleackley M.R, & Anderson M.A. (2020). Protein markers for Candida albicans EVs include claudin-like Sur7 family proteins. Journal of Extracellular Vesicles, 9(1), 1750810.
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