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Proceedings Paper

Whole-animal imaging with high spatio-temporal resolution
Author(s): Raghav Chhetri; Fernando Amat; Yinan Wan; Burkhard Höckendorf; William C. Lemon; Philipp J. Keller
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Paper Abstract

We developed isotropic multiview (IsoView) light-sheet microscopy in order to image fast cellular dynamics, such as cell movements in an entire developing embryo or neuronal activity throughput an entire brain or nervous system, with high resolution in all dimensions, high imaging speeds, good physical coverage and low photo-damage. To achieve high temporal resolution and high spatial resolution at the same time, IsoView microscopy rapidly images large specimens via simultaneous light-sheet illumination and fluorescence detection along four orthogonal directions. In a post-processing step, these four views are then combined by means of high-throughput multiview deconvolution to yield images with a system resolution of ≤ 450 nm in all three dimensions. Using IsoView microscopy, we performed whole-animal functional imaging of Drosophila embryos and larvae at a spatial resolution of 1.1-2.5 μm and at a temporal resolution of 2 Hz for up to 9 hours. We also performed whole-brain functional imaging in larval zebrafish and multicolor imaging of fast cellular dynamics across entire, gastrulating Drosophila embryos with isotropic, sub-cellular resolution. Compared with conventional (spatially anisotropic) 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, such as lattice lightsheet microscopy or diSPIM, IsoView microscopy effectively doubles the penetration depth and provides subsecond temporal resolution for specimens 400-fold larger than could previously be imaged.

Paper Details

Date Published: 7 March 2016
PDF: 6 pages
Proc. SPIE 9720, High-Speed Biomedical Imaging and Spectroscopy: Toward Big Data Instrumentation and Management, 97200R (7 March 2016); doi: 10.1117/12.2212564
Show Author Affiliations
Raghav Chhetri, Howard Hughes Medical Institute (United States)
Fernando Amat, Howard Hughes Medical Institute (United States)
Yinan Wan, Howard Hughes Medical Institute (United States)
Burkhard Höckendorf, Howard Hughes Medical Institute (United States)
William C. Lemon, Howard Hughes Medical Institute (United States)
Philipp J. Keller, Howard Hughes Medical Institute (United States)


Published in SPIE Proceedings Vol. 9720:
High-Speed Biomedical Imaging and Spectroscopy: Toward Big Data Instrumentation and Management
Kevin K. Tsia; Keisuke Goda, Editor(s)

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