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:

Nanoscale resolution in GFP-based microscopy

Nature Methods volume 3, pages 721–723 (2006)Cite this article

Abstract

We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy with GFP-labeled samples. The ∼70 nm lateral resolution attained in this study is demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell. Our results mark the advent of nanoscale biological microscopy with genetically encoded markers.

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: STED microscope for GFP imaging.
Figure 2: STED overcomes the diffraction resolution barrier in GFP-based microscopy.
Figure 3: Subdiffraction imaging of GFP-labeled ER in PtK2 cell (a–d) Confocal (a) and STED (b) images.

Similar content being viewed by others

References

  1. Pawley, J.B. (ed.) Handbook of biological confocal microscopy (Springer, New York, 2006).

    Book  Google Scholar 

  2. Conchello, J-A. & Lichtman, J.W. Nat. Methods 2, 920–931 (2005).

    Article  CAS  Google Scholar 

  3. Hell, S.W. & Wichmann, J. Opt. Lett. 19, 780–782 (1994).

    Article  CAS  Google Scholar 

  4. Klar, T.A., Jakobs, S., Dyba, M., Egner, A. & Hell, S.W. Proc. Natl. Acad. Sci. USA 97, 8206–8210 (2000).

    Article  CAS  Google Scholar 

  5. Hell, S.W. Nat. Biotechnol. 21, 1347–1355 (2003).

    Article  CAS  Google Scholar 

  6. Westphal, V. & Hell, S.W. Phys. Rev. Lett. 94, 143903 (2005).

    Article  Google Scholar 

  7. Willig, K.I., Rizzoli, S.O., Westphal, V., Jahn, R. & Hell, S.W. Nature 440, 935–939 (2006).

    Article  CAS  Google Scholar 

  8. Kittel, R.J. et al. Science 312, 1051–1054 (2006).

    Article  CAS  Google Scholar 

  9. Sieber, J.J., Willig, K.I., Heintzmann, R., Hell, S.W. & Lang, T. Biophys. J. 90, 2843–2851 (2006).

    Article  CAS  Google Scholar 

  10. Dyba, M., Jakobs, S. & Hell, S.W. Nat. Biotechnol. 21, 1303–1304 (2003).

    Article  CAS  Google Scholar 

  11. Schäfer, F.P. Dye Lasers (Springer, Berlin, 1973).

    Google Scholar 

  12. Charpilienne, A. J. Biol. Chem. 276, 29361–29367 (2001).

    Article  CAS  Google Scholar 

  13. Griesbeck, O., Baird, G.S., Campbell, R.E., Zacharias, D.A. & Tsien, R.Y. J. Biol. Chem. 276, 29188–29194 (2001).

    Article  CAS  Google Scholar 

  14. Campbell, R.E. et al. Proc. Natl. Acad. Sci. USA 99, 7877–7882 (2002).

    Article  CAS  Google Scholar 

  15. Shaner, N.C., Steinbach, P.A. & Tsien, R.Y. Nat. Methods 2, 905–909 (2005).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Purified GFP-VLPs were provided by J. Cohen and the plasmid eGFP-ER by P. Lipp. We thank S. Verrier, T. Rosenmund, A.C. Schauss, J.J. Sieber and T. Müller for providing samples, A. Schönle for help with the software ImSpector, V. Westphal and B. Harke for valuable discussions, as well as J. Keller and B. Rankin for critical reading. We thank R.Y. Tsien for providing the plasmids coding for mCitrine, mStrawbery and mRFP.

Author information

Authors and Affiliations

  1. Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, Göttingen, 37077, Germany

    Katrin I Willig, Robert R Kellner, Rebecca Medda, Birka Hein, Stefan Jakobs & Stefan W Hell

Authors
  1. Katrin I Willig
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Robert R Kellner
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rebecca Medda
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Birka Hein
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Stefan Jakobs
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Stefan W Hell
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Stefan W Hell.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Willig, K., Kellner, R., Medda, R. et al. Nanoscale resolution in GFP-based microscopy. Nat Methods 3, 721–723 (2006). https://doi.org/10.1038/nmeth922

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth922

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