PUBLICATION
Whole-animal functional and developmental imaging with isotropic spatial resolution
- Authors
- Chhetri, R.K., Amat, F., Wan, Y., Höckendorf, B., Lemon, W.C., Keller, P.J.
- ID
- ZDB-PUB-151027-2
- 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
Citation
Chhetri, R.K., Amat, F., Wan, Y., Höckendorf, B., Lemon, W.C., Keller, P.J. (2015) Whole-animal functional and developmental imaging with isotropic spatial resolution. Nature Methods. 12(12):1171-8.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping