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.

  • Correspondence
  • Published:

Live-cell dSTORM with SNAP-tag fusion proteins

Nature Methods volume 8, pages 7–9 (2011)Cite this article

Subjects

To the Editor:

In the September 2010 issue of Nature Methods we demonstrated live-cell direct stochastic optical reconstruction microscopy (dSTORM) of histone H2B proteins using a trimethoprim chemical tag (TMP tag) for genetic encoding with photostable standard fluorophores1. The method takes advantage of the fact that cells contain the reducing thiol glutathione—a cysteine-containing tripeptide—at millimolar concentrations, which enables reversible photoswitching of synthetic organic fluorophores2,3. The generality of the method can be easily understood considering that most Alexa Fluors (Invitrogen) and Atto fluorophores (ATTO-TEC) belong to the class of rhodamine and oxazine derivatives that have similar redox properties, that is, the triplet state of rhodamine and oxazine fluorophores is reduced by thiols such as glutathione3.

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

Relevant articles

Open Access articles citing this article.

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: Live-cell dSTORM with SNAP tags, and impact of direct excitation of radical anions at 405 nm.

References

  1. Wombacher, R. et al. Nat. Methods 7, 717–719 (2010).

    Article  CAS  Google Scholar 

  2. van de Linde, S. et al. Appl. Phys. B 93, 725–731 (2008).

    Article  CAS  Google Scholar 

  3. Heilemann, M. et al. Angew. Chem. Int. Edn. 48, 6903–6908 (2009).

    Article  CAS  Google Scholar 

  4. Keppler, A. et al. Nat. Biotechnol. 21, 86–89 (2003).

    Article  CAS  Google Scholar 

  5. van de Linde, S. et al. Photochem. Photobiol. Sci. published online 10 December 2010 (doi:10.1039/C0PP00317D).

    Article  CAS  Google Scholar 

  6. Betzig, E. et al. Science 313, 1642–1645 (2006).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank B. Vogel and T. Holm for assistance with experiments. This work was supported by the Biophotonics Initiative of the Bundesministerium für Bildung und Forschung.

Author information

Authors and Affiliations

  1. Department of Biotechnology and Biophysics, Julius Maximilians University Wuerzburg, Wuerzburg, Germany

    Teresa Klein, Anna Löschberger, Sven Proppert, Steve Wolter, Sebastian van de Linde & Markus Sauer

Authors
  1. Teresa Klein
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anna Löschberger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sven Proppert
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Steve Wolter
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sebastian van de Linde
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Markus Sauer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Markus Sauer.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Methods (PDF 650 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Klein, T., Löschberger, A., Proppert, S. et al. Live-cell dSTORM with SNAP-tag fusion proteins. Nat Methods 8, 7–9 (2011). https://doi.org/10.1038/nmeth0111-7b

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth0111-7b

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