研究细胞或组织内特殊基因表达的调控机理，分析单细胞内蛋白质和mRNA的特殊基因表现很有必要。原位杂交对于发现mRNA是必不可缺少的方法，它不但可以分析特殊mRNA的表达，而且可以证实特殊蛋白质的合成部位。原位杂交取决于细胞或组织内靶核酸与核酸探针的碱基配对，且条件尽可能优化。作为核酸探针，人工合成的01igo-DNA颇受青睐，且在诸多方面优于克隆双链CDNA。应用01igo-DNA探针可在组织切片上清楚地将28srRNA定位于细胞核或细胞质部位。原位杂交28srRNA对于评价切片中可杂交的rRNA水平十分方便。随着计算机图象分析广泛应用和酶免疫组织化学技术半抗原探针的问世，非放射性比色测定染色法也在所进步，借助显微照相与图象分析仪可以定量检测各种染色密度，或借助于扫描电镜技术背散电子探针以显示特殊的mRNA的位置。原位杂交非放射法最佳优点是它的高分辨率。这使人们可以在细胞和亚细胞水平分析mRNA的分布。事实上，应用此法研究mR－NA不规则分布方面已有明显进步。如将actinmRNA定位于细胞质；乙酸胆碱脂酶的mRNA仅限于核周区，而此种酶被输送到特定的细胞膜部位；胃泌素mRNA仅限于人胃泌素细胞的核上区，而翻译了的胃泌素蛋白质则被输送到细胞的基底部；雌激素受体mRNA的分布随雌激素治疗后猴子 It is necessary to study the regulation mechanism of special gene expression in cells or tissues and to analyze the special gene expression of proteins and mRNAs in single cells. In situ hybridization is essential for the discovery of mRNA, which not only analyzes the expression of specific mRNAs but also confirms the synthetic site of a particular protein. In situ hybridization is based on base pairing of the target nucleic acid to the nucleic acid probe within the cell or tissue, and the conditions are optimized as much as possible. Synthetic 01igo-DNA is favored as a nucleic acid probe and is superior to the cloned double-stranded CDNA in many aspects. Using the 01igo-DNA probe, 28srRNA can be clearly localized to the nucleus or cytoplasm on tissue sections. In situ hybridization 28srRNA is very convenient for assessing the level of hybridizable rRNA in sections. With the widespread use of computerized image analysis and the advent of hapten probes by enzyme immunohistochemistry, non-radioactive colorimetric staining has also been advanced, with the aid of micrographs and image analyzers for quantitative detection of various dye concentrations, Or backscatter the electron probe by means of scanning electron microscopy to reveal the location of the particular mRNA. The best advantage of non-radioactive in situ hybridization is its high resolution. This allows one to analyze the distribution of mRNAs at the cellular and subcellular level. In fact, significant improvements have been made in the study of the irregular distribution of mR-NA using this method. Such as actin mRNA localization in the cytoplasm; acetylcholine lipase mRNA only in the perinuclear zone, and this enzyme is transported to specific cell membrane sites; gastrin mRNA limited to human gastrin cells in the nuclear region, and the translation The gastrin protein is transported to the base of the cell; estrogen receptor mRNA distribution with estrogen-treated monkeys