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:

Modular scanning FCS quantifies receptor-ligand interactions in living multicellular organisms

Nature Methods volume 6, pages 643–645 (2009)Cite this article

Abstract

Analysis of receptor-ligand interactions in vivo is key to biology but poses a considerable challenge to quantitative microscopy. Here we combine static-volume, two-focus and dual-color scanning fluorescence correlation spectroscopy to solve this task at cellular resolution in complex biological environments. We quantified the mobility of fibroblast growth factor receptors Fgfr1 and Fgfr4 in cell membranes of living zebrafish embryos and determined their in vivo binding affinities to their ligand Fgf8.

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: Principle of scanning FCS.
Figure 2: Modular scanning FCS in cell membranes of living zebrafish embryos.

Similar content being viewed by others

References

  1. de Jong, L.A.A., Uges, D.R.A., Franke, J.P. & Bischoff, R. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 829, 1–25 (2005).

    Article  CAS  PubMed  Google Scholar 

  2. Ornitz, D.M. et al. J. Biol. Chem. 271, 15292–15297 (1996).

    Article  CAS  PubMed  Google Scholar 

  3. Olsen, S. et al. Genes Dev. 20, 185–198 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Yamaguchi, T.P., Harpal, K., Henkemeyer, M. & Rossant, J. Genes Dev. 8, 3032–3044 (1994).

    Article  CAS  PubMed  Google Scholar 

  5. Deng, C.X. et al. Genes Dev. 8, 3045–3057 (1994).

    Article  CAS  PubMed  Google Scholar 

  6. Weinstein, M., Xu, X., Ohyama, K. & Deng, C.X. Development 125, 3615–3623 (1998).

    CAS  PubMed  Google Scholar 

  7. Scholpp, S., Groth, C., Lohs, C., Lardelli, M. & Brand, M. Dev. Genes Evol. 214, 285–295 (2004).

    Article  CAS  PubMed  Google Scholar 

  8. Bacia, K., Kim, S. & Schwille, P. Nat. Methods 3, 83–89 (2006).

    Article  CAS  PubMed  Google Scholar 

  9. Yu, S.R. et al. Nature in the press.

  10. Ries, J. & Schwille, P. Phys. Chem. Chem. Phys. 10, 3487–3497 (2008).

    Article  CAS  PubMed  Google Scholar 

  11. Ries, J. & Schwille, P. Biophys. J. 91, 1915–1924 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Bottcher, R. & Niehrs, C. Endocr. Rev. 26, 63–77 (2005).

    Article  PubMed  Google Scholar 

  13. Reifers, F. et al. Development 125, 2381–2395 (1998).

    CAS  PubMed  Google Scholar 

  14. Scholpp, S. & Brand, M. Curr. Biol. 14, 1834–1841 (2004).

    Article  CAS  PubMed  Google Scholar 

  15. Hou, S., Maccarana, M., Min, T.H., Strate, I. & Pera, E.M. Dev. Cell 13, 226–241 (2007).

    Article  CAS  PubMed  Google Scholar 

  16. Maeder, C.I. et al. Nat. Cell Biol. 9, 1319–1326 (2007).

    Article  CAS  PubMed  Google Scholar 

  17. Magatti, D. & Ferri, F. Appl. Opt. 40, 4011–4021 (2001).

    Article  CAS  PubMed  Google Scholar 

  18. Sambrook, J. & Russell, D. Molecular Cloning: A Laboratory Manual (CSHL Press, Cold Spring Harbor, New York, USA, 2001).

    Google Scholar 

Download references

Acknowledgements

We thank E.P. Petrov, Z. Petrasek, M. Nowak and A. Picker for help writing the manuscript and E. Sieber for technical help. This work was supported by a Human Frontier Science Program network grant (050503-50) to M.B. and P.S. and by EU Integrated Project 'Zebrafish Models' to M.B.

Author information

Author notes

  1. Jonas Ries and Shuizi Rachel Yu: These authors contributed equally to this work.

Authors and Affiliations

  1. Biotechnology Center, Technical University of Dresden, Dresden, Germany

    Jonas Ries, Shuizi Rachel Yu, Markus Burkhardt, Michael Brand & Petra Schwille

  2. Center for Regenerative Therapies, Technical University of Dresden, Dresden, Germany

    Shuizi Rachel Yu, Michael Brand & Petra Schwille

Authors
  1. Jonas Ries
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shuizi Rachel Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Markus Burkhardt
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Michael Brand
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Petra Schwille
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Michael Brand or Petra Schwille.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Tables 1–2 and Supplementary Notes 1–4 (PDF 2165 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ries, J., Yu, S., Burkhardt, M. et al. Modular scanning FCS quantifies receptor-ligand interactions in living multicellular organisms. Nat Methods 6, 643–645 (2009). https://doi.org/10.1038/nmeth.1355

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.1355

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