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Controlling protein adsorption on graphene for cryo-EM using low-energy hydrogen plasmas

An Erratum to this article was published on 27 June 2014

This article has been updated

Abstract

Despite its many favorable properties as a sample support for biological electron microscopy, graphene is not widely used because its hydrophobicity precludes reliable protein deposition. We describe a method to modify graphene with a low-energy hydrogen plasma, which reduces hydrophobicity without degrading the graphene lattice. Use of plasma-treated graphene enables better control of protein distribution in ice for electron cryo-microscopy and improves image quality by reducing radiation-induced sample motion.

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Figure 1: Low-energy hydrogen-plasma treatment renders graphene hydrophilic and removes contamination.
Figure 2: Dose-dependent adsorption of proteins on hydrogen plasma–treated graphene.
Figure 3: Reduced motion of proteins on graphene substrates, as shown by speed plots.

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  • 10 June 2014

    In the version of this article initially published, the arrow in Figure 1b pointing to the 0–110 reflection was in the incorrect location. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank J.A. Golovchenko for the use of a chemical vapor–deposition instrument at Harvard for graphene synthesis during the initial phases of this work; S. Scotcher for fabrication of custom sample holders and copper punch machines; I.S. Fernandez, A. Kelley and V. Ramakrishnan of the Medical Research Council (MRC) Laboratory of Molecular Biology for the gift of ribosomes; J. Grimmett, T. Darling, G. McMullan and S. Chen for technical assistance; and E. Rajendra, R.A. Crowther, D. Neuhaus, X.C. Bai, S. Scheres, N. Unwin, A.R. Faruqi and R. Henderson for helpful discussions and comments. This work was supported by the European Research Council (ERC) under the European Union's Seventh Framework Programme (FP7/2007-2013)/ERC grant agreement no. 261151 to L.A.P., an MRC (UK) Centenary award to C.J.R. and MRC (UK) grant MC_U105192715 (L.A.P.).

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C.J.R. designed and performed the experiments and analyzed the data. C.J.R. and L.A.P. designed experiments, planned the project, interpreted the results and wrote the manuscript.

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Correspondence to Lori A Passmore.

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Competing interests

C.J.R. and L.A.P. are inventors on a patent application related to technology described in this manuscript (US 61/865,359).

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Supplementary Figures 1–9, Supplementary Table 1 and Supplementary Notes 1 and 2 (PDF 7263 kb)

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Russo, C., Passmore, L. Controlling protein adsorption on graphene for cryo-EM using low-energy hydrogen plasmas. Nat Methods 11, 649–652 (2014). https://doi.org/10.1038/nmeth.2931

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