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In-Situ Hybridization and Immunohistochemistry in Whole Embryos

  • Carol Irving
Part of the METHODS IN MOLECULAR BIOLOGY™ book series (MIMB, volume 461)

1. Introduction

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...

Keywords

Lithium Chloride Short Incubation Time Sheep Serum Concentrate Stock Solution Prehybridization Solution 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

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.

References

  1. 1.
    Wilkinson DG (1992) Whole mount in situ hybridization of vertebrate embryos. In: Wilkinson DG (ed) In situ hybridization. a practical approach. IRL Press, Oxford University Press, Oxford, UK.Google Scholar
  2. 2.
    Henrique D et al. (1995) Nature 375:787–790CrossRefPubMedGoogle Scholar
  3. 3.
    Hamburger V, Hamilton L (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92.CrossRefGoogle Scholar

Copyright information

© Humana Press, a Part of Springer Science + Business Media, LLC 2008

Authors and Affiliations

  • Carol Irving
    • 1
  1. 1.DePartment of Anatomy and Developmental BiologyUniversity CollegeLondonUK

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