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:

High-resolution three-dimensional imaging of large specimens with light sheet–based microscopy

Abstract

We report that single (or selective) plane illumination microscopy (SPIM), combined with a new deconvolution algorithm, provides a three-dimensional spatial resolution exceeding that of confocal fluorescence microscopy in large samples. We demonstrate this by imaging large living multicellular specimens obtained in a three-dimensional cell culture. The ability to rapidly image large samples at high resolution with minimal photodamage provides new opportunities especially for the study of subcellular processes in large living specimens.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1: Comparison of SPIM and confocal fluorescence microscopy by imaging the autofluorescence of grains of paper mulberry pollen.
Figure 2: Comparison of SPIM and confocal fluorescence microscopy of MDCK cysts.
Figure 3: Images recorded using SPIM of a large cellular spheroid of BxPC3 human pancreatic cancer cells labeled with DRAQ5.

Similar content being viewed by others

References

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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  3. Keller, P.J., Pampaloni, F. & Stelzer, E.H.K. Curr. Opin. Cell Biol. 18, 117–124 (2006).

    Article  CAS  Google Scholar 

  4. Greger, K., Swoger, J. & Stelzer, E.H.K. Rev. Sci. Instrum. (in the press).

  5. Swoger, J., Huisken, J. & Stelzer, E.H.K. Opt. Lett. 28, 1654–1656 (2003).

    Article  Google Scholar 

  6. Agard, D.A. & Sedat, J.W. Nature 302, 676–681 (1983).

    Article  CAS  Google Scholar 

  7. Carrington, W.A. et al. Science 268, 1483–1487 (1995).

    Article  CAS  Google Scholar 

  8. Verveer, P.J. & Jovin, T.M. Appl. Opt. 37, 6240–6246 (1998).

    Article  CAS  Google Scholar 

  9. Timmins, N.E., Dietmair, S. & Nielsen, L.K. Angiogenesis 7, 97–103 (2004).

    Article  Google Scholar 

  10. Kunz-Schughart, L.A., Freyer, J.P., Hofstaedter, F. & Ebner, R. J. Biomol. Screen. 9, 273–285 (2004).

    Article  CAS  Google Scholar 

  11. Kam, Z., Hanser, B., Gustafsson, M.G., Agard, D.A. & Sedat, J.W. Proc. Natl. Acad. Sci. USA 98, 3790–3795 (2001).

    Article  CAS  Google Scholar 

  12. Engelbrecht, C.J. & Stelzer, E.H.K. Opt. Lett. 31, 1477–1479 (2006).

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank D. Holzer (European Molecular Biology Laboratory, Heidelberg) for help with MDCK cell culture and transfection, and A. Schrödel and M. Löhr (German Cancer Research Center, Heidelberg) for providing the BxPC3 spheroids. E.H.K.S. and F.P. acknowledge the Forschungsprogramm Optische Technologien der Landesstiftung Baden-Württemberg for financial support. M.M. was supported by a joint collaboration between Hamamatsu and the German Cancer Research Center (Project PA 11631).

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Peter J Verveer, Jim Swoger or Ernst H K Stelzer.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

SPIM and confocal microscopy of CHO cells. (PDF 141 kb)

Supplementary Fig. 2

SPIM deconvolution of a Medaka embryo. (PDF 148 kb)

Supplementary Fig. 3

Confocal microscopy of pancreatic tumor cells. (PDF 91 kb)

Supplementary Methods (PDF 114 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Verveer, P., Swoger, J., Pampaloni, F. et al. High-resolution three-dimensional imaging of large specimens with light sheet–based microscopy. Nat Methods 4, 311–313 (2007). https://doi.org/10.1038/nmeth1017

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth1017

This article is cited by

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