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      A guide to light-sheet fluorescence microscopy for multiscale imaging

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      Journal: Nature Methods
      Publisher: Springer Nature

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          Abstract

          This Review introduces the fundamental considerations for building a light sheet microscope, describes the pros and cons associated with available implementations, and offers practical advice for users.

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          Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy.

          Philipp Keller, Annette Schmidt, Joachim Wittbrodt … (2008)
          A long-standing goal of biology is to map the behavior of all cells during vertebrate embryogenesis. We developed digital scanned laser light sheet fluorescence microscopy and recorded nuclei localization and movement in entire wild-type and mutant zebrafish embryos over the first 24 hours of development. Multiview in vivo imaging at 1.5 billion voxels per minute provides "digital embryos," that is, comprehensive databases of cell positions, divisions, and migratory tracks. Our analysis of global cell division patterns reveals a maternally defined initial morphodynamic symmetry break, which identifies the embryonic body axis. We further derive a model of germ layer formation and show that the mesendoderm forms from one-third of the embryo's cells in a single event. Our digital embryos, with 55 million nucleus entries, are provided as a resource.
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            Lattice light-sheet microscopy: imaging molecules to embryos at high spatiotemporal resolution.

            Bi-Chang Chen, Wesley R Legant, Kai Wang … (2014)
            Although fluorescence microscopy provides a crucial window into the physiology of living specimens, many biological processes are too fragile, are too small, or occur too rapidly to see clearly with existing tools. We crafted ultrathin light sheets from two-dimensional optical lattices that allowed us to image three-dimensional (3D) dynamics for hundreds of volumes, often at subsecond intervals, at the diffraction limit and beyond. We applied this to systems spanning four orders of magnitude in space and time, including the diffusion of single transcription factor molecules in stem cell spheroids, the dynamic instability of mitotic microtubules, the immunological synapse, neutrophil motility in a 3D matrix, and embryogenesis in Caenorhabditis elegans and Drosophila melanogaster. The results provide a visceral reminder of the beauty and the complexity of living systems. Copyright © 2014, American Association for the Advancement of Science.
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              Optical sectioning deep inside live embryos by selective plane illumination microscopy.

              Jan Huisken, Jim Swoger, Filippo Del Bene … (2004)
              Large, living biological specimens present challenges to existing optical imaging techniques because of their absorptive and scattering properties. We developed selective plane illumination microscopy (SPIM) to generate multidimensional images of samples up to a few millimeters in size. The system combines two-dimensional illumination with orthogonal camera-based detection to achieve high-resolution, optically sectioned imaging throughout the sample, with minimal photodamage and at speeds capable of capturing transient biological phenomena. We used SPIM to visualize all muscles in vivo in the transgenic Medaka line Arnie, which expresses green fluorescent protein in muscle tissue. We also demonstrate that SPIM can be applied to visualize the embryogenesis of the relatively opaque Drosophila melanogaster in vivo.
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                Author and article information

                Journal
                Title: Nature Methods
                Abbreviated Title: Nat Meth
                Publisher: Springer Nature
                ISSN (Print): 1548-7091
                ISSN (Electronic): 1548-7105
                Publication date Created: March 31 2017
                Publication date (Print): March 31 2017
                Volume: 14
                Issue: 4
                Pages: 360-373
                Article
                DOI: 10.1038/nmeth.4224
                SO-VID: 8620c803-0674-413c-b7b4-9b2fd14d8bb7
                Copyright © © 2017
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