原位杂交技术在培养细胞冰冻切片和石蜡片切中的应用

ESTABLISHMENT OF IN SITU HYBRIDIZATION METHOD AN ITS APPLICATION TO CULTURED CELLS, FROZEN SECTIONS AND PARAFFIN EMBEDDED SECTIONS

本实验详细地探讨了原位杂交技术的有关问题:生物素标记DNA探针的制备,各类来源标本的制备,杂交前、中、后细胞组织切片的处理等;并把缺口翻译法合成的生物素化EB病毒DNA探针应用于检测培养细胞,冰冻和石蜡切片中EB病毒Bam HI-W重复片断DNA序列。实验结果表明:自制的EB病毒DNA探针完全可靠;所采用的一套原位杂交技术适用于检测上述三类不同的标本,其检测的结果是满意的。

In situ hybridization method, Which has been developed on the basis of liquid-phase hybridization, has great sensitivity and high accuracy. It will certainly provide important investigative and diagnostic tool for pathologic study. The authors have explored in de-tail of establishing in situr hybridization method in this experiment: preparation of biotiny-lated hybridization probe, preparation of different sorts of sample, treatment of cells and tissue sections in pre-hybridization, hybridization and post-hybridization, and so on. We also applied the biotinylated EBV-DNA probe which was prepared by means of nick-translation to cultured cells, frozen sections and paraffin embedded sections for examining EBV-DNA Bam HI-W internal repeat fragment. The findings showed that the EBV-DNA probe prepared by us is completely reliable, the in situ hybridization procedure can be applied to the above three sorts of sample and the results are satisfactory.