Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

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

Nature Methods volume 11, pages 649–652 (2014)Cite this article

Subjects

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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

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.

Similar content being viewed by others

Accession codes

Accessions

Electron Microscopy Data Bank

Protein Data Bank

Change history

  • 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.

References

  1. Grigorieff, N. & Harrison, S.C. Curr. Opin. Struct. Biol. 21, 265–273 (2011).

    Article  CAS  Google Scholar 

  2. Bai, X.-C., Fernandez, I.S., McMullan, G. & Scheres, S.H. Elife 2, e00461 (2013).

    Article  Google Scholar 

  3. Dubochet, J. et al. Q. Rev. Biophys. 21, 129–228 (1988).

    Article  CAS  Google Scholar 

  4. Robertson, J. Adv. Phys. 35, 317–374 (1986).

    Article  CAS  Google Scholar 

  5. Miyazawa, A. et al. J. Mol. Biol. 288, 765–786 (1999).

    Article  CAS  Google Scholar 

  6. Brilot, A.F. et al. J. Struct. Biol. 177, 630–637 (2012).

    Article  CAS  Google Scholar 

  7. Henderson, R. & McMullan, G. Microscopy 62, 43–50 (2013).

    Article  Google Scholar 

  8. Geim, A.K. Science 324, 1530–1534 (2009).

    Article  CAS  Google Scholar 

  9. Lee, C., Wei, X., Kysar, J.W. & Hone, J. Science 321, 385–388 (2008).

    Article  CAS  Google Scholar 

  10. Russo, C.J. A structural imaging study of single DNA molecules on carbon nanotubes. PhD thesis, Harvard Univ. (2010).

  11. Pantelic, R.S., Meyer, J.C., Kaiser, U., Baumeister, W. & Plitzko, J.M. J. Struct. Biol. 170, 152–156 (2010).

    Article  CAS  Google Scholar 

  12. Pantelic, R.S. et al. J. Struct. Biol. 174, 234–238 (2011).

    Article  CAS  Google Scholar 

  13. Sader, K. et al. J. Struct. Biol. 183, 531–536 (2013).

    Article  CAS  Google Scholar 

  14. Gómez-Navarro, C. et al. Nano Lett. 7, 3499–3503 (2007).

    Article  Google Scholar 

  15. Elias, D.C. et al. Science 323, 610–613 (2009).

    Article  CAS  Google Scholar 

  16. Méndez, I. et al. J. Phys. Chem. A 110, 6060–6066 (2006).

    Article  Google Scholar 

  17. Russo, C.J. & Golovchenko, J.A. Proc. Natl. Acad. Sci. USA 109, 5953–5957 (2012).

    Article  CAS  Google Scholar 

  18. Israelachvili, J. & Pashley, R. Nature 300, 341–342 (1982).

    Article  CAS  Google Scholar 

  19. Curtis, G.H. & Ferrier, R.P. J. Phys. D Appl. Phys. 2, 1035–1040 (1969).

    Article  Google Scholar 

  20. Downing, K.H., McCartney, M.R. & Glaeser, R.M. Microsc. Microanal. 10, 783–789 (2004).

    Article  CAS  Google Scholar 

  21. Reina, A. et al. Nano Lett. 9, 30–35 (2009).

    Article  CAS  Google Scholar 

  22. Li, X. et al. Science 324, 1312–1314 (2009).

    Article  CAS  Google Scholar 

  23. Lieberman, M.A. & Lichtenberg, A.J. Principles of Plasma Discharges and Materials Processing 2nd edn. (Wiley, 2005).

  24. Taherian, F., Marcon, V., van der Vegt, N.F.A. & Leroy, F. Langmuir 29, 1457–1465 (2013).

    Article  CAS  Google Scholar 

  25. Dubochet, J., Groom, M. & Mueller-Neuteboom, S. Adv. Opt. Electron Microsc. 8, 107–135 (1982).

    CAS  Google Scholar 

  26. Regan, W. et al. Appl. Phys. Lett. 96, 113102 (2010).

    Article  Google Scholar 

  27. Selmer, M. et al. Science 313, 1935–1942 (2006).

    Article  CAS  Google Scholar 

  28. Wasilewski, S., Karelina, D., Berriman, J.A. & Rosenthal, P.B. J. Struct. Biol. 180, 243–248 (2012).

    Article  CAS  Google Scholar 

  29. Tang, G. et al. J. Struct. Biol. 157, 38–46 (2007).

    Article  CAS  Google Scholar 

  30. Scheres, S.H.W. J. Struct. Biol. 180, 519–530 (2012).

    Article  CAS  Google Scholar 

  31. Mindell, J.A. & Grigorieff, N. J. Struct. Biol. 142, 334–347 (2003).

    Article  Google Scholar 

  32. Voorhees, R.M., Weixlbaumer, A., Loakes, D., Kelley, A.C. & Ramakrishnan, V. Nat. Struct. Mol. Biol. 16, 528–533 (2009).

    Article  CAS  Google Scholar 

  33. Pettersen, E.F. et al. J. Comput. Chem. 25, 1605–1612 (2004).

    Article  CAS  Google Scholar 

  34. Armache, J.-P. et al. Proc. Natl. Acad. Sci. USA 107, 19754–19759 (2010).

    Article  CAS  Google Scholar 

  35. Scheres, S.H.W. & Chen, S. Nat. Methods 9, 853–854 (2012).

    Article  CAS  Google Scholar 

  36. Rosenthal, P.B. & Henderson, R. J. Mol. Biol. 333, 721–745 (2003).

    Article  CAS  Google Scholar 

Download references

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.).

Author information

Authors and Affiliations

  1. Medical Research Council Laboratory of Molecular Biology, Cambridge, UK

    Christopher J Russo & Lori A Passmore

Authors
  1. Christopher J Russo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lori A Passmore
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

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.

Corresponding author

Correspondence to Lori A Passmore.

Ethics declarations

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).

Supplementary information

Supplementary Text and Figures

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

Source data

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.2931

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing