Whole-animal functional and developmental imaging with isotropic spatial resolution

Imaging fast cellular dynamics across large specimens requires high resolution in all dimensions, high imaging speeds, good physical coverage, and low photodamage. To meet these requirements, we developed isotropic multiview (IsoView) light-sheet microscopy, which rapidly images large specimens via simultaneous light-sheet illumination and fluorescence detection along with four orthogonal directions. Combining these four views using high-throughput multiview deconvolution yields images with high resolution in all three dimensions. We demonstrate whole-animal functional imaging of Drosophila larvae at a spatial resolution of 1.1-2.5 μm and temporal resolution of 2 Hz for several hours. We also present spatially isotropic whole-brain functional imaging in Danio rerio larvae and spatially isotropic multicolor imaging of fast cellular dynamics across gastrulating Drosophila embryos. IsoView microscopy improves spatial resolution at least sevenfold and decreases resolution anisotropy at least threefold compared with conventional light-sheet microscopy. In addition, IsoView microscopy effectively doubles the penetration depth and provides subsecond temporal resolution for specimens 400-fold larger than could previously be imaged compared with existing high-resolution light-sheet techniques.

The publication of the IsoView microscope is available from the literature section (Chhetri, Amat, Wan, Höckendorf, Lemon and Keller 2015, Nature Methods).

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