An optimized procedure for whole-mount in situ hybridization on mouse embryos and embryoid bodies.
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.
Piette
Denis
D
Vrije Universiteit Brussel (VUB), Department of Biology, Lab for Cell Genetics, Pleinlaan 2, 1050 Brussels, Belgium.
Hendrickx
Marijke
M
Willems
Erik
E
Kemp
Caroline R
CR
Leyns
Luc
L
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
D018075:RNA, Complementary