title: An optimized procedure for whole-mount in situ hybridization on mouse embryos and embryoid bodies.
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.
authors.affiliation: Vrije Universiteit Brussel (VUB), Department of Biology, Lab for Cell Genetics, Pleinlaan 2, 1050 Brussels, Belgium.
authors.forename: Denis
authors.initials: D
authors.lastname: Piette
authors.forename: Marijke
authors.initials: M
authors.lastname: Hendrickx
authors.forename: Erik
authors.initials: E
authors.lastname: Willems
authors.forename: Caroline R
authors.initials: CR
authors.lastname: Kemp
authors.forename: Luc
authors.initials: L
authors.lastname: Leyns
mesh_terms: D000818:Animals; D004622:Embryo, Mammalian / Q000378:metabolism*; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods*; D017403:In Situ Hybridization / Q000379:methods*; D051379:Mice; D018075:RNA, Complementary / Q000235:genetics
chemical_list: D018075:RNA, Complementary