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. 2015 Dec;12(12):1171-8.
doi: 10.1038/nmeth.3632. Epub 2015 Oct 26.

Whole-animal functional and developmental imaging with isotropic spatial resolution

Raghav K Chhetri  1 , Fernando Amat  1 , Yinan Wan  1 , Burkhard Höckendorf  1 , William C Lemon  1 , Philipp J Keller  1
Affiliations

Whole-animal functional and developmental imaging with isotropic spatial resolution

Raghav K Chhetri et al. Nat Methods. 2015 Dec.

Abstract

Imaging fast cellular dynamics across large specimens requires high resolution in all dimensions, high imaging speeds, good physical coverage and low photo-damage. 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 four orthogonal directions. Combining these four views by means of 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. Compared with conventional light-sheet microscopy, IsoView microscopy improves spatial resolution at least sevenfold and decreases resolution anisotropy at least threefold. Compared with existing high-resolution light-sheet techniques, IsoView microscopy effectively doubles the penetration depth and provides subsecond temporal resolution for specimens 400-fold larger than could previously be imaged.

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References

    1. Nat Methods. 2015 May;12(5):379-80 - PubMed
    1. Nat Methods. 2012 Jun 03;9(7):755-63 - PubMed
    1. Science. 2004 Aug 13;305(5686):1007-9 - PubMed
    1. Opt Express. 2002 Jan 28;10(2):145-54 - PubMed
    1. Nat Biotechnol. 2013 Nov;31(11):1032-8 - PubMed

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