A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:00Title
- 01:00Introduction
- 01:27Fixing and Dechorionating Embryos
- 03:10Permeabilizing the embryos
- 03:51Hybridization
- 05:37Antibody labeling
- 09:10Propidium iodide staining and detection
- 10:31Outcome and Results
- 11:41Conclusion
Whole mount in situ hybridization is one of the most widely used techniques in developmental biology. Here, we present a high-resolution double fluorescent in situ hybridization protocol for analyzing the precise expression pattern of a single gene and for determining the overlap of the expression domains of two genes. We include a propidium iodide nuclear counter-stain to highlight tissue organization.
Tags
ZebrafishWhole MountHigh-resolutionDouble FluorescentIn Situ HybridizationDevelopmental BiologyGene ExpressionGene OverlapAlkaline PhosphataseNBT/BCIPFast RedDigoxygeninFluorescein Labeled ProbesTyramide Signal Amplification (TSA)Nuclear StainsPropidium IodideFluorescence ImmunohistochemistryReactive Fluorescent SubstrateTyrosine ResiduesAntisense RiboprobeStaining PatternsLaser Scanning Confocal MicroscopyNascent TranscriptsChromosomal LociNuclear StainingCytoplasmic Staining
