Rumballe Bree A, Chiu Han Sheng, Georgas Kylie M, Little Melissa H
Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, Australia.
Methods Mol Biol. 2012;886:223-39. doi: 10.1007/978-1-61779-851-1_20.
Studies into the molecular basis of morphogenesis frequently begin with investigations into gene expression across time and cell type in that organ. One of the most anatomically informative approaches to such studies is the use of in situ hybridization, either of intact or histologically sectioned tissues. Here, we describe the optimization of this approach for use in the temporal and spatial analysis of gene expression in the urogenital system, from embryonic development to the postnatal period. The methods described are applicable for high throughput analysis of large gene sets. As such, ISH has become a powerful technique for gene expression profiling and is valuable for the validation of profiling analyses performed using other approaches such as microarrays.
对形态发生分子基础的研究通常始于对该器官中跨时间和细胞类型的基因表达进行调查。此类研究中最具解剖学信息价值的方法之一是使用原位杂交,可用于完整组织或经组织学切片的组织。在此,我们描述了这种方法的优化,以用于泌尿生殖系统中从胚胎发育到出生后阶段基因表达的时空分析。所描述的方法适用于对大型基因集进行高通量分析。因此,原位杂交已成为一种用于基因表达谱分析的强大技术,对于验证使用其他方法(如微阵列)进行的谱分析非常有价值。