PUBLICATION

Imaging and 3D Reconstruction of Cerebrovascular Structures in Embryonic Zebrafish

Authors
Ethell, D.W., Cameron, D.J.
ID
ZDB-PUB-140513-39
Date
2014
Source
Journal of visualized experiments : JoVE   (86): (Journal)
Registered Authors
Keywords
none
MeSH Terms
  • Animals
  • Brain/blood supply*
  • Brain/embryology*
  • Cerebrovascular Circulation
  • Female
  • Green Fluorescent Proteins/chemistry
  • Imaging, Three-Dimensional/methods*
  • Male
  • Zebrafish/embryology*
PubMed
24797110 Full text @ J. Vis. Exp.
Abstract
Zebrafish are a powerful tool to study developmental biology and pathology in vivo. The small size and relative transparency of zebrafish embryos make them particularly useful for the visual examination of processes such as heart and vascular development. In several recent studies transgenic zebrafish that express EGFP in vascular endothelial cells were used to image and analyze complex vascular networks in the brain and retina, using confocal microscopy. Descriptions are provided to prepare, treat and image zebrafish embryos that express enhanced green fluorescent protein (EGFP), and then generate comprehensive 3D renderings of the cerebrovascular system. Protocols include the treatment of embryos, confocal imaging, and fixation protocols that preserve EGFP fluorescence. Further, useful tips on obtaining high-quality images of cerebrovascular structures, such as removal the eye without damaging nearby neural tissue are provided. Potential pitfalls with confocal imaging are discussed, along with the steps necessary to generate 3D reconstructions from confocal image stacks using freely available open source software.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping