Combined FISH and immunofluorescent staining methods to co-localize proteins and mRNA in neurons and brain tissue.
Combined FISH and immunofluorescent staining methods to co-localize proteins and mRNA in neurons and brain tissue.
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DOI：
10.1007/978-1-62703-411-1_9
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发表时间：
2013-01-01
期刊：
影响因子：
--
通讯作者：
Tanese, Naoko
中科院分区：
文献类型：
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作者：
Ma, Bin;Tanese, Naoko
多色荧光原位杂交(FISH)和免疫荧光染色(IFS)相结合，提供了一种显示不同神经室中mRNA和蛋白质之间的空间关系的有效方法。尽管表面上看起来简单明了，但将IFS/FISH与定量的信使核糖核酸和蛋白质共定位分析结合起来，可能很难成功进行，往往会产生不同的结果。在这里，我们描述了一种结合二维(2D)和三维(3D)共定位分析的多色迭代函数系统和FISH方法来确定单个分子在大鼠神经元和脑切片中的表达。用这种方法，我们已经分析了亨廷顿病蛋白Huntingtin与选定的蛋白质和mRNA的相互作用。
Combining multicolor fluorescent in situ hybridization (FISH) and immunofluorescent staining (IFS) presents a powerful method for visualizing the spatial relationship between mRNA and proteins in different neural compartments. Although seemingly straightforward, the combination of IFS/FISH and quantitative co-localization analysis of mRNA and proteins can be difficult to perform successfully, often generating variable results. Here we describe a combined method of multicolor IFS and FISH in concert with two-dimensional (2D) and three-dimensional (3D) co-localization analysis for determining the expression of individual molecules in rat neurons and brain sections. Using this approach, we have analyzed interactions of the Huntington's disease protein huntingtin with select proteins and mRNA.