Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan.
Center for Highly Advanced Integration of Nano and Life Sciences (G-CHAIN), Gifu University, Gifu, Gifu, Japan.
Methods Mol Biol. 2022;2303:719-730. doi: 10.1007/978-1-0716-1398-6_54.
In situ hybridization provides information for understanding the localization of gene expression in various tissues. The relative expression levels of mRNAs in a single cell can be sensitively visualized by this technique. Furthermore, since in situ hybridization is a histological technique, tissue structure is maintained after fixation, and it is possible to accurately identify the cell types. We have examined the expression of heparan sulfate sulfotransferases by in situ hybridization to better understand the functions of heparan sulfate in the development of mouse nervous system. This chapter describes methods of in situ hybridization analyses using cRNA probes labeled with non-radioactive nucleotides.
原位杂交为理解基因在各种组织中的表达定位提供了信息。该技术可以灵敏地显示单个细胞中 mRNA 的相对表达水平。此外,由于原位杂交是一种组织学技术,在固定后保留了组织结构,因此可以准确识别细胞类型。我们通过原位杂交检查了硫酸乙酰肝素硫酸转移酶的表达,以更好地理解硫酸乙酰肝素在小鼠神经系统发育中的功能。本章描述了使用非放射性核苷酸标记的 cRNA 探针进行原位杂交分析的方法。