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.

  • Commentary
  • Published:

Light-sheet fluorescence microscopy for quantitative biology

Nature Methods volume 12, pages 23–26 (2015)Cite this article

Subjects

In light sheet–based fluorescence microscopy (LSFM), optical sectioning in the excitation process minimizes fluorophore bleaching and phototoxic effects. Because biological specimens survive long-term three-dimensional imaging at high spatiotemporal resolution, LSFM has become the tool of choice in developmental biology.

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: Basic optical arrangement in LSFM.
Figure 2: LSFM imaging of a live Tribolium castaneum embryo expressing nuclear GFP.

References

  1. Cox, I.J. J. Microsc. 133, 149–154 (1984).

    Article  CAS  Google Scholar 

  2. Wijnaendts Van Resandt, R.W. et al. J. Microsc. 138, 29–34 (1985).

    Article  Google Scholar 

  3. Wagner, M. et al. Int. J. Mol. Sci. 11, 956–966 (2010).

    Article  CAS  Google Scholar 

  4. Huisken, J., Swoger, J., Del Bene, F., Wittbrodt, J. & Stelzer, E.H.K. Science 305, 1007–1009 (2004).

    Article  CAS  Google Scholar 

  5. Greger, K., Swoger, J. & Stelzer, E.H.K. Rev. Sci. Instrum. 78, 023705 (2007).

    Article  CAS  Google Scholar 

  6. Keller, P.J., Schmidt, A.D., Wittbrodt, J. & Stelzer, E.H.K. Science 322, 1065–1069 (2008).

    Article  CAS  Google Scholar 

  7. Wilson, T. & Sheppard, C.J.R. Theory and Practice of Scanning Optical Microscopy (Academic Press, 1984).

    Google Scholar 

  8. Breuninger, T., Greger, K. & Stelzer, E.H.K. Opt. Lett. 32, 1938–1940 (2007).

    Article  Google Scholar 

  9. Keller, P.J. et al. Nat. Methods 7, 637–642 (2010).

    Article  CAS  Google Scholar 

  10. Truong, T.V., Supatto, W., Koos, D.S., Choi, J.M. & Fraser, S.E. Nat. Methods 8, 757–760 (2011).

    Article  CAS  Google Scholar 

  11. Fahrbach, F.O. & Rohrbach, A. Opt. Express 18, 24229–24244 (2010).

    Article  Google Scholar 

  12. Vettenburg, T. et al. Nat. Methods 11, 541–544 (2014).

    Article  CAS  Google Scholar 

  13. Cella Zanacchi, F. et al. Nat. Methods 8, 1047–1049 (2011).

    Article  Google Scholar 

  14. Huang, B., Wang, W., Bates, M. & Zhuang, X. Science 319, 810–813 (2008).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  16. Chen, B.-C. et al. Science 346, 1257998 (2014).

    Article  Google Scholar 

  17. Gustafsson, M.G.L. J. Microsc. 198, 82–87 (2000).

    Article  CAS  Google Scholar 

  18. Siedentopf, H. & Zsigmondy, R. Ann. Phys. 315, 1–39 (1903).

    Article  Google Scholar 

  19. Voie, A.H., Burns, D.H. & Spelman, F.A. J. Microsc. 170, 229–236 (1993).

    Article  CAS  Google Scholar 

  20. Verveer, P.J. et al. Nat. Methods 4, 311–313 (2007).

    Article  CAS  Google Scholar 

  21. Swoger, J., Verveer, P., Greger, K., Huisken, J. & Stelzer, E.H.K. Opt. Express 15, 8029–8042 (2007).

    Article  Google Scholar 

  22. Hell, S. & Stelzer, E.H.K. J. Opt. Soc. Am. A 9, 2159–2166 (1992).

    Article  Google Scholar 

  23. Stelzer, E.H.K. & Lindek, S. Opt. Commun. 111, 536–547 (1994).

    Article  Google Scholar 

  24. Strobl, F. & Stelzer, E.H.K. Development 141, 2331–2338 (2014).

    Article  CAS  Google Scholar 

  25. Maizel, A., von Wangenheim, D., Federici, F., Haseloff, J. & Stelzer, E.H.K. Plant J. 68, 377–385 (2011).

    Article  CAS  Google Scholar 

  26. Wohland, T., Shi, X.K., Sankaran, J. & Stelzer, E.H.K. Opt. Express 18, 10627–10641 (2010).

    Article  CAS  Google Scholar 

  27. Greger, K., Neetz, M.J., Reynaud, E.G. & Stelzer, E.H.K. Opt. Express 19, 20743–20750 (2011).

    Article  CAS  Google Scholar 

  28. Preibisch, S. et al. Nat. Methods 11, 645–648 (2014).

    Article  CAS  Google Scholar 

  29. Wu, Y. et al. Proc. Natl. Acad. Sci. USA 108, 17708–17713 (2011).

    Article  CAS  Google Scholar 

  30. Pampaloni, F. et al. Integr. Biol. (Camb.) 6, 988–998 (2014).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

E.H.K.S.'s research is funded through CEF-MC (EXC 115) by the Deutsche Forschungsgemeinschaft.

Author information

Authors and Affiliations

  1. Ernst H.K. Stelzer is at the Buchmann Institute for Molecular Life Sciences, Fachbereich Lebenswissenschaften (FB15, IZN), Goethe Universität Frankfurt am Main, Frankfurt am Main, Germany.,

    Ernst H K Stelzer

Authors
  1. Ernst H K Stelzer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ernst H K Stelzer.

Ethics declarations

Competing interests

E.H.K.S. is an inventor or co-inventor of two relevant patents owned by the European Molecular Biology Laboratory Enterprise Management Technology Transfer GmbH: (i) microscope with a viewing direction perpendicular to the illumination direction, US 7554725; DE 10257423 and (ii) single plane illumination microscope US 20070109633 A1; PCT/EP03/05991.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Stelzer, E. Light-sheet fluorescence microscopy for quantitative biology. Nat Methods 12, 23–26 (2015). https://doi.org/10.1038/nmeth.3219

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.3219

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