ZFIN ID: ZDB-PUB-151027-2
Whole-animal functional and developmental imaging with isotropic spatial resolution
Chhetri, R.K., Amat, F., Wan, Y., Höckendorf, B., Lemon, W.C., Keller, P.J.
Date: 2015
Source: Nature Methods 12(12): 1171-8 (Journal)
Registered Authors: Amat, Fernando, Keller, Philipp, Lemon, William, Wan, Yinan
Keywords: none
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
PubMed: 26501515 Full text @ Nat. Methods
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.