摘要
彗星试验是近十年发展起来的在单细胞水平上检测ＤＮＡ断裂的方法，理论上可检测含有大量ＤＮＡ碎片的凋亡细胞。本文在建立地塞米松诱导小鼠胸腺细胞凋亡模型的基础上，探索了彗星试验检测凋亡的实验方法，并与Ｇｉｅｍｓａ染色、琼脂糖凝胶电泳、ＴＵＮＥＬ法等几种常用的检测凋亡细胞的方法进行了比较研究。结果表明：凋亡细胞在彗星试验中呈现特征性的彗星形态，很小的彗星头，大而圆的彗星尾；以彗星试验检测细胞凋亡的最适电泳条件为２０Ｖ，１５ｍｉｎ；其检测灵敏度与Ｇｉｅｍｓａ染色法和ＴＵＮＥＬ法相当。因此彗星试验是一种灵敏、准确、简便、既可定性又可定量检测细胞凋亡的新方法。
Comet assay is a method measuring DNA strand breaks on single cell level, which has been developed for about 10 years. Theoretically, apoptotic cells with extensive DNA internucleosomal fragments can be detected by this method. On the basis that apoptosis in immature mice thymocytes could be induced by dexamethasone(DEX), the detection of apoptotic cells by comet assay was studied and compared with agarose gel electrophoresis, morphological observations with Giemsa staining and TUNEL assay. Apoptotic cells showed a comet feature with a very small head, and a large and wide tail. The optimal conditions for electrophoresis in comet assay was 20 volt in 15 minutes. Comet assay is sensitive, valid, rapid and simple and is highly correlated with Giemsa staining and TUNEL assay in detecting apoptotic cells.
出处
《卫生研究》
CAS
CSCD
北大核心
1999年第2期83-85,共3页
Journal of Hygiene Research
基金
国家自然科学基金
