Large-scale whole mount in situ hybridization of mouse embryos.
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.
Shimizu
Hirohito
H
Department of Systems Biomedicine, National Research Institute for Child Health and Development, Tokyo, Japan.
Uchibe
Kenta
K
Asahara
Hiroshi
H
D000818:Animals; D030541:Databases, Genetic; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods; D017403:In Situ Hybridization / Q000295:instrumentation / Q000379:methods*; D051379:Mice; D008967:Molecular Biology / Q000379:methods; D011247:Pregnancy; D015347:RNA Probes
D015347:RNA Probes