PUBLICATION
Labeling and Imaging Cells in the Zebrafish Hindbrain
- Authors
- Jayachandran, P., Hong, E., and Brewster, R.
- ID
- ZDB-PUB-100811-3
- Date
- 2010
- Source
- Journal of visualized experiments : JoVE (41): (Journal)
- Registered Authors
- Brewster, Rachel, Hong, Elim
- Keywords
- none
- MeSH Terms
-
- Animals
- DNA/administration & dosage
- DNA/genetics
- Embryo, Nonmammalian/cytology
- Green Fluorescent Proteins/analysis*
- Green Fluorescent Proteins/biosynthesis
- Green Fluorescent Proteins/genetics
- Microinjections/methods
- Microscopy, Confocal/methods*
- Plasmids/genetics
- Rhombencephalon/cytology
- Rhombencephalon/embryology*
- Zebrafish/embryology*
- PubMed
- 20689510 Full text @ J. Vis. Exp.
Citation
Jayachandran, P., Hong, E., and Brewster, R. (2010) Labeling and Imaging Cells in the Zebrafish Hindbrain. Journal of visualized experiments : JoVE. (41).
Abstract
Key to understanding the morphogenetic processes that shape the early vertebrate embryo is the ability to image cells at high resolution. In zebrafish embryos, injection of plasmid DNA results in mosaic expression, allowing for the visualization of single cells or small clusters of cells (1) . We describe how injection of plasmid DNA encoding membrane-targeted Green Fluorescent Protein (mGFP) under the control of a ubiquitous promoter can be used for imaging cells undergoing neurulation. Central to this protocol is the methodology for imaging labeled cells at high resolution in sections and also in real time. This protocol entails the injection of mGFP DNA into young zebrafish embryos. Embryos are then processed for vibratome sectioning, antibody labeling and imaging with a confocal microscope. Alternatively, live embryos expressing mGFP can be imaged using time-lapse confocal microscopy. We have previously used this straightforward approach to analyze the cellular behaviors that drive neural tube formation in the hindbrain region of zebrafish embryos (2). The fixed preparations allowed for unprecedented visualization of cell shapes and organization in the neural tube while live imaging complemented this approach enabling a better understanding of the cellular dynamics that take place during neurulation.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping