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Guide to light-sheet microscopy for adventurous biologists

Nature Methods volume 12, pages 30–34 (2015)Cite this article

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Ten years of development in light-sheet microscopy have led to spectacular demonstrations of its capabilities. The technology is ready to assist biologists in tackling scientific problems, but are biologists ready for it? Here we discuss the interdisciplinary challenges light-sheet microscopy presents for biologists and highlight available resources.

Light-sheet microscopy has the unique ability to image biological systems for extended periods of time and in three dimensions without damaging the specimens. This ability is in large part due to the uncoupling of the illumination and detection axes of the microscope, so that only the part of the specimen that is imaged gets illuminated. Additionally, the detection of fluorescent signals with a wide-field detection device such as a charge-coupled device (CCD) camera allows stunning imaging speeds. Together these properties open up windows into biological processes at unprecedented spatiotemporal scales, but as usual, there is no free lunch. The impressive capabilities of light-sheet microscopes come at a price: they demand a level of interdisciplinarity previously unseen in the biological sciences.

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Figure 1: Current light-sheet microscopes.
Figure 2: (a) 3D rendering of the OpenSPIM design.
Figure 3: The Big Data Challenge.

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Acknowledgements

We thank all members of the ever growing light-sheet community who push this field forward. All participants, speakers and instructors of the first EMBO practical course on light-sheet microscopy in Dresden inspired us to get this piece written. We thank J. Oegema for Figure 3b and L. Gutierrez-Heredia for Figure 2b.

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Authors and Affiliations

  1. Emmanuel G. Reynaud is at the Earth Institute, University College Dublin, Dublin, Ireland.,

    Emmanuel G Reynaud

  2. Jan Peychl, Jan Huisken and Pavel Tomancak are at the Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.,

    Jan Peychl, Jan Huisken & Pavel Tomancak

Authors
  1. Emmanuel G Reynaud
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  2. Jan Peychl
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  3. Jan Huisken
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  4. Pavel Tomancak
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Correspondence to Pavel Tomancak.

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Reynaud, E., Peychl, J., Huisken, J. et al. Guide to light-sheet microscopy for adventurous biologists. Nat Methods 12, 30–34 (2015). https://doi.org/10.1038/nmeth.3222

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