title: Rapid and stable buffer exchange system using InSitu Chip suitable for multicolor and large-scale whole-mount analyses.
abstract: Whole-mount in situ hybridization (WISH) and whole-mount immunohistochemistry (WIHC) are informative methods commonly used to analyze the spatiotemporal and quantitative distribution of mRNAs and proteins. However, these methods require multiple buffer changes and the imposition of time- and nerve-consuming efforts. To facilitate the whole-mount analyses, we innovated an easy and one-step buffer exchange system named "InSitu Chip" based on a single column containing two attached filters. This system improves the speed and stabilizes the different steps of the currently available protocols, providing fast and uniform operations. The InSitu Chip system is especially appropriate for multicolor whole-mount analyses using fluorescent detection. Furthermore, the InSitu Chip system is also suitable for large-scale whole-mount experiments associated with genome, transcriptome, and/or proteome analyses requiring high-throughput, high-quality, and reproducible results. Using the InSitu Chip, about 1,500 gene expression patterns were stably surveyed in ascidian Ciona intestinalis juveniles.
authors.affiliation: Department of Biology, Faculty of Science, Chiba University, Inage-ku, Japan. ogasawara@faculty.chiba-u.jp
authors.forename: Michio
authors.initials: M
authors.lastname: Ogasawara
authors.forename: Nori
authors.initials: N
authors.lastname: Satoh
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Shimada
authors.forename: Zhipeng
authors.initials: Z
authors.lastname: Wang
authors.forename: Toshio
authors.initials: T
authors.lastname: Tanaka
authors.forename: Sumihare
authors.initials: S
authors.lastname: Noji
mesh_terms: D000818:Animals; D002021:Buffers; D002938:Ciona intestinalis / Q000235:genetics; D020869:Gene Expression Profiling / Q000295:instrumentation*; D007150:Immunohistochemistry / Q000295:instrumentation*; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation*; D040901:Proteomics / Q000295:instrumentation*
chemical_list: D002021:Buffers