石蜡包埋组织上进行染色体特异性DNA探针原位杂交的可行性

Arnoldus E P, Dreef E J, Noordermeer I A, Verheggen M M, Thierry R F, Peters A C, Cornelisse C J, Van der Ploeg M, Raap A K

Department of Cytochemistry and Cytometry, Leiden University, The Netherlands.

J Clin Pathol. 1991 Nov;44(11):900-4. doi: 10.1136/jcp.44.11.900.

The feasibility was studied of in situ hybridisation using chromosome specific DNA probes on paraffin wax embedded normal and malignant tissues from different organs. Both isolated nuclei and 5 microns sections were used in in situ hybridisation experiments with biotinylated repetitive DNA probes specific for the centromeric regions of chromosomes 1 and 17. The hybridisation results were visualised with peroxidase-diaminobenzidine. The optimal pretreatments with sodium thiocyanate and pepsin were experimentally defined for the different tissues. Although interphase cytogenetics on paraffin wax embedded tissue is possible, the results indicate that it has its limitations, compared with investigations on fresh tumour tissue.

研究了使用染色体特异性DNA探针，对来自不同器官的石蜡包埋正常组织和恶性组织进行原位杂交的可行性。在原位杂交实验中，针对1号和17号染色体着丝粒区域的生物素化重复DNA探针，使用了分离的细胞核和5微米切片。杂交结果用过氧化物酶-二氨基联苯胺进行可视化。通过实验确定了不同组织用硫氰酸钠和胃蛋白酶的最佳预处理方法。虽然对石蜡包埋组织进行间期细胞遗传学分析是可行的，但结果表明，与对新鲜肿瘤组织的研究相比，它有其局限性。