Feldmann G, Bernuau D
INSERM U24, Faculté de Médecine Xavier-Bichat, Paris.
Bull Assoc Anat (Nancy). 1989 Mar;73(220):17-26.
In situ hybridization is a molecular biological technique recently introduced in histology. Its principle consists of forming stable nucleic hybrids in tissues or cells. In this review, the main steps of the technique are discussed in regard to the preparation of probes and their labelling, the fixation of tissues and cells and their permeabilization in order to facilitate the penetration of labelled probes. Conditions of hybridization and requisite control reactions are also analysed. Quantification possible when radioactive probes are used is detailed. The three current applications of in situ hybridization, the localization of a gene on a chromosome, the demonstration of viral genomes in cells and the investigation of messenger RNAs coding for a determined protein, are illustrated with specific examples.
原位杂交是组织学中最近引入的一种分子生物学技术。其原理是在组织或细胞中形成稳定的核酸杂交体。在这篇综述中,将从探针的制备及其标记、组织和细胞的固定及其通透化等方面讨论该技术的主要步骤,以便于标记探针的穿透。还分析了杂交条件和必要的对照反应。详细介绍了使用放射性探针时可能进行的定量分析。文中通过具体实例说明了原位杂交目前的三种应用,即基因在染色体上的定位、细胞中病毒基因组的显示以及对编码特定蛋白质的信使核糖核酸的研究。