In-Situ Hybridization and Immunohistochemistry in Whole Embryos
The ability to visualize zones of active gene expression in whole embryos or tissues is a powerful tool toward understanding spatial and temporal relationships among molecules during development. Combining simultaneously a number of RNA probes with immunohistochemical detection of proteins allows direct comparison between transcription and translation of any given gene, a direct spatiotemporal comparison of a number of molecules, or can be used to identify exactly their expression domain within the embryo with relation to stained “tissue landmarks.”
Performing in-situ hybridization and immunohistochemistry before sectioning is advantageous, as larger sample numbers can be processed with speed. Sites of specific interest can subsequently be sectioned for visualization at a single-cell resolution.
A number of protocols are currently available, each with modifications for different organisms. Here, we describe a protocol that can be used for a variety of species and is...
KeywordsLithium Chloride Short Incubation Time Sheep Serum Concentrate Stock Solution Prehybridization Solution
I thank colleagues and members of the laboratory who contributed to the development and revision of these protocols. Work in the author's laboratory is supported by the MRC and the Royal Society.
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