整胚原位杂交技术是利用反义RNA探针检测体内mRNA表达的一项技术, 在利用模式动物研究基因时空表达方面有着重要的应用。如何使用该技术得到特异、高敏感度的表达结果, 对每一个使用该技术的实验室来说都很重要。本实验室参照常规的实验方法, 对该技术加以改进, 使之更加灵敏, 结果更加特异。文章主要以斑马鱼为例, 介绍了整胚原位杂交技术的发展历史, 并重点介绍了本实验室所用的整胚原位杂交实验流程, 同时还分析了实验结果不理想的原因及其解决方法。
Whole mount in situ hybridization (WISH) is a popular technique that is used to study temporal-spatial gene expression at mRNA level in model systems. Sensitive and high-resolution WISH is important for every laboratory that uses this technique to study expression pattern of genes in wildtype and perturbed embryos. Based on the commonly used proto-cols in fish community, our lab optimizes them to get more sensitive and specific results. Here, we briefly summarize the history of this technology using zebrafish as an example, describe a detailed experimental protocol, and provide trouble-shooting for imperfect results and their solution.
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