Abstract
Determining the precise expression pattern of a gene of interest at various stages of development is essential to understanding its biological function in embryology. This protocol describes a sensitive method for whole-mount in situ hybridization (WISH) to mouse embryos, using cRNA probes. Adaptations are provided that allow the protocol to be applied to embryonic stages ranging from blastocysts to postimplantation stage embryos, and to embryoid bodies. We also describe an in situ method for differential detection of two probes. Probe labeling and dissection and preparation of the embryos can be performed in 2 d. The actual WISH procedure can be completed in another 3 d.
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Acknowledgements
We thank Ann Pauwels and Ellen Geeroms for their help. The laboratory is supported by the Horizontal Action and Concerted Research Action of the Vrije Universiteit Brussel (Grants HOA1 and GOA29), the Flemish Fund for Scientific Research (Grants G.0485.06 and G.0307.04) and the EU (6th Framework, Grant NEST012930). M.H. is an aspirant FWO.
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Piette, D., Hendrickx, M., Willems, E. et al. An optimized procedure for whole-mount in situ hybridization on mouse embryos and embryoid bodies. Nat Protoc 3, 1194–1201 (2008). https://doi.org/10.1038/nprot.2008.103
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DOI: https://doi.org/10.1038/nprot.2008.103
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