彗星试验银染法的优化及验证  

Improvement of the silver-stain and it's test and verification in gomet assay

作　　者：赵蓉[1] 衡正昌[1] 

机构地区：[1]四川大学华西公共卫生学院环境卫生教研室,成都610041

出　　处：《卫生研究》2003年第2期107-109,共3页Journal of Hygiene Research

基　　金：国家自然科学基金资助项目 (No .39670 62 5)

摘　　要：在彗星试验中寻找一种最佳染色效果的银染法 ,拟取代溴化乙啶 (EB)染色。对选出的银染法从固定、染色、显色、脱色、染色温度等各个方面进行多次试验 ,达到最佳染色效果后 ,将优化的银染法用于检测两类化合物 -重铬酸钾 (K2 Cr2 O7)和 1,2—二氯乙烷 (1,2—dichloroethane,DCE)对两种类型细胞—小鼠脾细胞、人外周血淋巴细胞的DNA损伤作用 ,并与EB染色比较灵敏度。结果显示优化后的银染法是在彗星试验中成功建立的非荧光染色法 ,该法染色清晰、持久、背景颜色浅、染色对比度好、染色效果满意 ,方法操作简便、价廉 ;该法可检测出化合物对细胞的DNA损伤作用 ,与EB染色有相同阳性检出阈 ,可染出EB染色无法染上的DNA小片段。In order to find out a better silver stain method so that to replace EB stain in comet assay, the selected silver stain methods were tested via determining fixing, coloring, displaying, fading, and testing the pigmentation temperature. When it was optimized, it was used in testing the DNA damage by two types of compounds, namely potassium dichromate (K 2Cr 2O 7 ) and 1,2 dichloroethane( DCE ) to two kind of cells, namely spleen cells of mouse and human outer lymphocyte, and, the sensitivity was compared with that of EB stain. The results showed that the modified silver stain was the first successful non fluorescent dye method in comet assay that can dye clearly, permanently, with a light background and good contrast, and it was easy to manipulate and it was cheap. The stain can detect the DNA damage by compounds to cells and has the same positive threshold like that in EB stain. It can even dye the small fragments that EB stain cannot. It is suggested that in comet assay, the silver stain can substitute the EB stain completely.

关 键 词：彗星试验 非荧光染色 银染法 

分 类 号：Q－336[生物学]