Summary
Whole-mount in situ hybridization (WISH) is a method to visualize gene expression within a whole organism. Although it is an essential approach in developmental biology to describe the gene expression patterns during embryogenesis, the simultaneous processing of numerous embryos in a large-scale experiment such as a screening assay has been difficult for an individual due to the limited capacity of the experimental setting. This chapter describes the optimal protocol for a WISH assay in a relatively large experimental scale, which allows for more efficient processing of embryos. The major improvement in the efficiency has been achieved by the several refinements such as the introduction of a quick method for probe synthesis, the use of mesh-buckets to facilitate handling samples, and the simplification of the conventional procedures. In combination with publicly accessible gene expression databases, WISH assays will be more favorable for a large-scale assay using mouse embryos.
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Acknowledgments
The authors are grateful to Shigetoshi Yokoyama for professional and valuable advice. This work was supported by Genome Network Project 2004–2008 by MEXT, and the Grant of Research on Child Health and Development by Ministry of Health, Labor, and Welfare.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Shimizu, H., Uchibe, K., Asahara, H. (2009). Large-Scale Whole Mount In Situ Hybridization of Mouse Embryos. In: Koga, H. (eds) Reverse Chemical Genetics. Methods in Molecular Biology™, vol 577. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-232-2_13
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DOI: https://doi.org/10.1007/978-1-60761-232-2_13
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Publisher Name: Humana Press, Totowa, NJ
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