Rapid and stable buffer exchange system using InSitu Chip suitable for multicolor and large-scale whole-mount analyses
Rapid and stable buffer exchange system using InSitu Chip suitable for multicolor and large-scale whole-mount analyses
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DOI：
10.1007/s00427-005-0031-x
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发表时间：
2006-02-01
影响因子：
2.4
通讯作者：
Noji, S
中科院分区：
文献类型：
--
作者：
Ogasawara, M;Satoh, N;Noji, S
整体原位杂交（WISH）和整体免疫组化（WIHC）是常用于分析mRNA和蛋白质时空和定量分布的信息方法。然而，这些方法需要多个缓冲区的变化和强加的时间和神经消耗的努力。为了便于整体分析，我们创新了一种简单的一步缓冲液交换系统，称为“原位芯片”，该系统基于含有两个附加过滤器的单个柱。该系统提高了速度并稳定了当前可用协议的不同步骤，提供了快速和统一的操作。InSitu芯片系统特别适用于使用荧光检测的微生物学整体包埋分析。此外，InSitu Chip系统还适用于与基因组、转录组和/或蛋白质组分析相关的大规模整体实验，这些实验需要高通量、高质量和可重现的结果。使用原位芯片，大约1,500个基因表达模式稳定调查海鞘海鞘青少年。
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.