title: Whole-mount in situ hybridization (WISH) optimized for gene expression analysis in mouse embryos and embryoid bodies.
abstract: Whole-mount in situ hybridization (WISH) is a technique widely used in developmental biology to study the localization of RNA sequences in intact tissues or whole organisms. In this chapter we present a detailed protocol that was optimized for gene expression analysis in early stage mouse embryos (5.5-10.5 days post-coitum) and embryoid bodies formed by differentiating embryonic stem cells and can be used for the detection of up to two distinct RNA sequences simultaneously. The initial steps of the procedure are the generation of the labeled riboprobe(s) and the embryo or embryoid body preparation, which can be completed in less than 2 days. The actual WISH procedure, comprised of the hybridization, the post-hybridization washes, and the immunological staining, can be completed in 3 days.
authors.affiliation: Department of Biology, Lab of Cell Genetics, Vrije Universiteit Brussel, Pleinlaan, 2, 1050, Brussels, Belgium, edakou@vub.ac.be.
authors.forename: Eleni
authors.initials: E
authors.lastname: Dakou
authors.forename: Nele
authors.initials: N
authors.lastname: Vanbekbergen
authors.forename: Sara
authors.initials: S
authors.lastname: Corradi
authors.forename: Caroline R
authors.initials: CR
authors.lastname: Kemp
authors.forename: Erik
authors.initials: E
authors.lastname: Willems
authors.forename: Luc
authors.initials: L
authors.lastname: Leyns
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D004622:Embryo, Mammalian / Q000378:metabolism* / Q000648:ultrastructure; D058732:Embryoid Bodies / Q000378:metabolism* / Q000648:ultrastructure; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization / Q000379:methods*; D051379:Mice / Q000196:embryology* / Q000235:genetics; D008867:Microtomy / Q000379:methods; D012313:RNA / Q000032:analysis* / Q000235:genetics; D016610:Tissue Embedding / Q000379:methods
chemical_list: D012313:RNA