小鼠组织的高通量石蜡切片原位杂交和双重免疫组织化学
High-throughput paraffin section in situ hybridization and dual immunohistochemistry on mouse tissues.
作者信息
Rumballe Bree, Georgas Kylie, Little Melissa H
机构信息
Institute for Molecular Bioscience, University of Queensland, St. Lucia, 4072, Australia.
出版信息
CSH Protoc. 2008 Jul 1;2008:pdb.prot5030. doi: 10.1101/pdb.prot5030.
Abstract
INTRODUCTIONSection in situ hybridization (SISH) is a high-resolution tool used to analyze gene expression patterns. This protocol utilizes the Tecan Freedom EVO150 platform to perform high-throughput SISH on paraffin sections to detect mRNA with a digoxigenin (DIG)-labeled probe. The slide is mounted and imaged before performing immunohistochemistry (IHC) on the same section. The dual reaction enables a marker of protein expression to be localized on the same section as the mRNA and facilitates more accurate annotation of the gene expression.
摘要
引言
原位杂交（SISH）是一种用于分析基因表达模式的高分辨率工具。本方案利用Tecan Freedom EVO150平台在石蜡切片上进行高通量SISH，以使用地高辛（DIG）标记的探针检测mRNA。在对同一切片进行免疫组织化学（IHC）之前，先对玻片进行固定和成像。这种双重反应能够将蛋白质表达标记定位在与mRNA相同的切片上，有助于更准确地注释基因表达。
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