One FISH, dFISH, Three FISH: Sensitive Methods of Whole-Mount Fluorescent In Situ Hybridization in Freshwater Planarians
One FISH, dFISH, Three FISH: Sensitive Methods of Whole-Mount Fluorescent In Situ Hybridization in Freshwater Planarians
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DOI：
10.1007/978-1-4939-2303-8_7
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发表时间：
2015-01-01
期刊：
影响因子：
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通讯作者：
Pearson, Bret J.
中科院分区：
文献类型：
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作者：
Brown, David D. R.;Pearson, Bret J.
作为淡水真涡虫（扁形虫）进一步发展为一个模型系统，最有价值的工具仍然是原位杂交（ISH）分析基因表达在整个安装成年动物（WISH）。在用于比色、单色检测的标准协议的优化期间，已经克服了多个障碍。然而，对涡虫的基因功能研究已经发展到几乎所有的研究都需要同时分析多个RNA的地步。因此，整个真涡虫群体已经付出了相当大的努力来创造实现这一目标的适当方法。在这里，我们总结了该领域，方法的演变，并描述了一种新的方法，更快速，更灵敏地检测多种RNA的多荧光ISH。
As freshwater planarians (flatworms) are further developed as a model system, the most valuable tool continues to be in situ hybridization (ISH) analysis of gene expression in whole mount adult animals (WISH). Multiple hurdles have been overcome during the optimization of a standard protocol for colorimetric, single color detections. However, gene function studies on planarians have evolved to the point where virtually all investigations require the analysis of multiple RNAs, simultaneously. Thus, considerable effort by the entire planarian community has been put forth to create adequate methodologies towards this goal. Here we summarize the field, methodological evolution, and describe a new method for more rapid and more sensitive detection of multiple RNAs by multi-fluorescent ISH.