PUBLICATION

Adaptive light-sheet microscopy for long-term, high-resolution imaging in living organisms

Authors
Royer, L.A., Lemon, W.C., Chhetri, R.K., Wan, Y., Coleman, M., Myers, E.W., Keller, P.J.
ID
ZDB-PUB-161101-1
Date
2016
Source
Nature Biotechnology   34(12): 1267-1278 (Journal)
Registered Authors
Keller, Philipp, Lemon, William, Royer, Loic, Wan, Yinan
Keywords
Embryogenesis, Fluorescence imaging, Light-sheet microscopy, Neuroscience
MeSH Terms
  • Algorithms*
  • Animals
  • Drosophila
  • Embryo, Nonmammalian/cytology*
  • Equipment Design
  • Equipment Failure Analysis
  • Feedback
  • Image Enhancement/instrumentation*
  • Image Enhancement/methods*
  • Lasers
  • Lenses
  • Lighting/instrumentation
  • Lighting/methods
  • Longitudinal Studies
  • Microscopy, Fluorescence/instrumentation*
  • Microscopy, Fluorescence/methods*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Zebrafish
PubMed
27798562 Full text @ Nat Biotechnol.
Abstract
Optimal image quality in light-sheet microscopy requires a perfect overlap between the illuminating light sheet and the focal plane of the detection objective. However, mismatches between the light-sheet and detection planes are common owing to the spatiotemporally varying optical properties of living specimens. Here we present the AutoPilot framework, an automated method for spatiotemporally adaptive imaging that integrates (i) a multi-view light-sheet microscope capable of digitally translating and rotating light-sheet and detection planes in three dimensions and (ii) a computational method that continuously optimizes spatial resolution across the specimen volume in real time. We demonstrate long-term adaptive imaging of entire developing zebrafish (Danio rerio) and Drosophila melanogaster embryos and perform adaptive whole-brain functional imaging in larval zebrafish. Our method improves spatial resolution and signal strength two to five-fold, recovers cellular and sub-cellular structures in many regions that are not resolved by non-adaptive imaging, adapts to spatiotemporal dynamics of genetically encoded fluorescent markers and robustly optimizes imaging performance during large-scale morphogenetic changes in living organisms.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping