Whole-animal functional and developmental imaging with isotropic spatial resolution

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / embryology
  • Brain / ultrastructure*
  • Drosophila / embryology
  • Embryo, Nonmammalian / physiology
  • Embryo, Nonmammalian / ultrastructure*
  • Embryonic Development
  • Equipment Design
  • Image Processing, Computer-Assisted / instrumentation
  • Image Processing, Computer-Assisted / methods*
  • Larva
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Whole Body Imaging / instrumentation
  • Whole Body Imaging / methods*
  • Zebrafish / embryology