胃癌组织甲基绿染色DNA/RNA图像分析

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（５７８％），而ＭＧ＿Ｐ染色为１７／２６（６５４％），两者符合率为８８２％。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０９９。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。

Quantification of DNA/RNA in gastric cancer tissue with lmage cytometry using methyl green_pyronin stain

AIMS To explore the reliability for the simultaneous quantitative assessment of both DNA and RNA on tissue sections using MG_P stain. METHODS The adjacent sections of 26 gastric cancers, which had been analysised for DNA pliodies by Feulgen stain, were stained with the standardized MG_P procedure. The sections were evaluated for the uptake of MG and pyronin by cell nuclei, nucleoli and cytoplasm using ICM. Cell nuclei were stained bright green with a clearly visible chromation texture by methyl green, while nucleoli and cytoplasm were stained red by pyronin. The sections were then destained and a Feugen reaction was performed. RESULTS The aneuploidy of the gastric cancer studied by the MG_P method was 17/26(65 4%), while that by Feulgen stain was 15/26(57 8%). The coefficient of correlation of the IOD between methyl green and Feulgen was 0 99 by the destaining method. CONCLUSIONS The standardized methyl green_pyronin stain is reliable for the simultaneous quantitative assessment of both RNA and DNA.