PUBLICATION

Fast imaging of live organisms with sculpted light sheets

Authors
Chmielewski, A.K., Kyrsting, A., Mahou, P., Wayland, M.T., Muresan, L., Evers, J.F., Kaminski, C.F.
ID
ZDB-PUB-170214-192
Date
2015
Source
Scientific Reports   5: 9385 (Journal)
Registered Authors
Keywords
Developmental biology, Light-sheet microscopy
MeSH Terms
  • Animals
  • Cell Nucleus/metabolism
  • Cell Nucleus/pathology
  • Embryo, Nonmammalian/metabolism
  • Embryo, Nonmammalian/pathology
  • Image Processing, Computer-Assisted
  • Imaging, Three-Dimensional
  • Microscopy, Fluorescence/instrumentation
  • Microscopy, Fluorescence/methods*
  • Zebrafish
PubMed
25893952 Full text @ Sci. Rep.
Abstract
Light-sheet microscopy is an increasingly popular technique in the life sciences due to its fast 3D imaging capability of fluorescent samples with low photo toxicity compared to confocal methods. In this work we present a new, fast, flexible and simple to implement method to optimize the illumination light-sheet to the requirement at hand. A telescope composed of two electrically tuneable lenses enables us to define thickness and position of the light-sheet independently but accurately within milliseconds, and therefore optimize image quality of the features of interest interactively. We demonstrated the practical benefit of this technique by 1) assembling large field of views from tiled single exposure each with individually optimized illumination settings; 2) sculpting the light-sheet to trace complex sample shapes within single exposures. This technique proved compatible with confocal line scanning detection, further improving image contrast and resolution. Finally, we determined the effect of light-sheet optimization in the context of scattering tissue, devising procedures for balancing image quality, field of view and acquisition speed.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping