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Improving your four-dimensional image: traveling through a decade of light-sheet-based fluorescence microscopy research

Nature Protocols volume 12, pages 1103–1109 (2017)Cite this article

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Abstract

Light-sheet-based fluorescence microscopy features optical sectioning in the excitation process. This reduces phototoxicity and photobleaching by up to four orders of magnitude compared with that caused by confocal fluorescence microscopy, simplifies segmentation and quantification for three-dimensional cell biology, and supports the transition from on-demand to systematic data acquisition in developmental biology applications.

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Figure 1: LSFM in three-dimensional cell biology.
Figure 2: LSFM in insect developmental biology.

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Acknowledgements

We thank K. Hötte (Goethe Universität, Frankfurt am Main, Germany) for the T47D spheroid images, S. Fischer for helpful comments on the manuscript, M.F. Schetelig (Justus-Liebig-Universität, Gießen, Germany) for the Ceratitis line, and T. Mito (Tokushima University, Tokushima, Japan) for the Gryllus line. The research was supported by funding from the Cluster of Excellence–Frankfurt am Main for Macromolecular Complexes (CEF-MC II, EXC 115; speaker: V. Dötsch) granted in part to E.H.K.S. at the Buchmann Institute for Molecular Life Sciences (BMLS; director: E. Schleiff) at the Johann Wolfgang Goethe Universität–Frankfurt am Main by the Deutsche Forschungsgemeinschaft (DFG).

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Authors and Affiliations

  1. Physical Biology/Physikalische Biologie (IZN, FB 15), Buchmann Institute for Molecular Life Sciences (BMLS), Cluster of Excellence Frankfurt–Macromolecular Complexes (CEF–MC), Goethe Universität–Frankfurt am Main (Campus Riedberg), Frankfurt am Main, Germany

    Frederic Strobl, Alexander Schmitz & Ernst H K Stelzer

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  1. Frederic Strobl
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F.S., A.S., and E.H.K.S. wrote the manuscript. F.S. and A.S. prepared the display items.

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Correspondence to Ernst H K Stelzer.

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Strobl, F., Schmitz, A. & Stelzer, E. Improving your four-dimensional image: traveling through a decade of light-sheet-based fluorescence microscopy research. Nat Protoc 12, 1103–1109 (2017). https://doi.org/10.1038/nprot.2017.028

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