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目的研究甲基绿＿派洛宁（ＭＧ＿Ｐ）染色法进行胃癌细胞的ＤＮＡ／ＲＮＡ双参数图像分析的可行性。方法选择２６例已用Ｆｅｕｌｇｅｎ染色法检测过ＤＮＡ倍体类型的胃癌石蜡组织，采用同一蜡块的连续切片进行ＭＧ＿Ｐ染色。提纯甲基绿并经反复摸索，找到了ＭＧ＿Ｐ染色的理想条件和最佳染色效果。细胞的染色质呈蓝绿色，核仁和胞浆呈红色，两种颜色对比明显；其他细胞结构不着色。经ＤＮＡ酶处理后染色质不着色，ＲＮＡ酶处理后核仁及胞浆不着色。用图像分析法同时测定胃癌细胞的ＤＮＡ／ＲＮＡ含量，并进行ＤＮＡ倍体分析。结果２６例胃癌患者Ｆｅｕｇｅｎ染色的异倍体率为１５／２６（５７８％），而ＭＧ＿Ｐ染色为１７／２６（６５４％），两者符合率为８８２％。采用退色的方法测定相同组织细胞的ＤＮＡ含量，Ｆｅｕｌｇｅｎ染色与ＭＧ＿Ｐ染色的相关系数的０９９。ＲＮＡ含量在肿瘤细胞中明显高于正常胃粘膜细胞。结论ＭＧ＿Ｐ染色法是可靠的同时测定肿瘤细胞ＤＮＡ／ＲＮＡ含量的方法。
Objective To investigate the feasibility of DNA/RNA dual-parameter image analysis of gastric cancer cells by methyl green-pyronin (MG_P) staining method. Methods Twenty-six paraffin-embedded gastric cancer tissues that had been tested for DNA ploidy by Feulgen staining were stained with MG_P in serial sections of the same paraffin block. The methyl green was purified and repeatedly explored to find the ideal conditions for MG_P staining and the best staining results. The chromatin of the cells is blue-green, and the nucleolus and cytoplasm are red. The contrast between the two colors is obvious; other cell structures are not colored. After DNase treatment, the chromatin is not stained, and after RNase treatment, the nucleolus and cytoplasm are not stained. Simultaneous determination of DNA/RNA content in gastric cancer cells by image analysis and DNA ploidy analysis. Results The aneuploidy rate of Feugen staining in 26 cases of gastric cancer was 15/26 (57.8%), while that of MG_P was 17/26 (65.4%). The coincidence rate was 88. 2%. The discoloration method was used to determine the DNA content of the same tissue cells, and the correlation coefficient of the Feulgen staining with MG_P staining was 099. The RNA content was significantly higher in tumor cells than in normal gastric mucosa cells. Conclusion MG_P staining is a reliable method for simultaneous determination of DNA/RNA content in tumor cells.