高内涵筛选结合活细胞实时成像技术分析谷氨酸诱导的小鼠神经瘤母细胞N2a凋亡  

Analysis of glutamate-induced N2a cell apoptosis using high-content screening and live cell real-time imaging

作　　者：乔可[1] 崔士超[2] 胡捷先 陈献华[2] 

机构地区：[1]复旦大学基础医学院医学分子病毒学教育部/卫生部重点实验室,上海200032 [2]复旦大学基础医学院医学分子病毒学教育部医学神经生物学国家重点实验室,上海200032

出　　处：《复旦学报（医学版）》2015年第4期538-542,共5页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金面上项目(31370788);国家自然科学基金重点项目(81030020)~~

摘　　要：目的利用高内涵筛选(high-content screening,HCS)技术结合活细胞实时成像技术研究谷氨酸诱导的小鼠神经瘤母细胞N2a凋亡的形态学变化特征。方法用HCS技术分析摸索不同浓度谷氨酸诱导的N2a细胞早期凋亡百分比的变化,同时结合活细胞实时成像技术追踪谷氨酸诱导的N2a凋亡的形态学变化特征。结果当谷氨酸浓度为750μg/mL时,发生早期凋亡的N2a细胞比例达到峰值(42.07%)。对同一谷氨酸浓度,随着拍摄追踪时间的增加,凋亡细胞数目逐渐增多,胞核荧光强度的变异系数逐渐增大,胞核面积逐渐缩小,胞核逐渐出现碎片化。结论使用HCS结合活细胞实时成像技术能直观清晰地捕捉谷氨酸诱导N2a细胞凋亡的形态学变化特征,是一种体外追踪单一细胞凋亡形态学变化的可靠方法。该方法也可应用于药物对细胞凋亡的促进或抑制作用的形态学变化特征的研究。Objective To analyze glutamate-induced mouse neuroblastoma N2a cell apoptosis using high-content screening and live cell real-time imaging. Methods The percentage of early apoptotic N2a cells induced by different concentration of glutamate was analyzed by high-content screening （HCS）. The characteristics of morphological variation of apoptotic N2a cells were tracked by live cell real-time imaging. Results The percentage of early apoptotic N2a cells reached the highest （42.07%） at the glutamate concentration of 750 /μg/mL. With the increase of tracking time, the statistic data for both the number of apoptotic N2a cells and the coefficient of variation （CV） for nucleus fluorescence intensity were increased, the nucleus areas was decreased as well, which was consistent with the apoptotic progress of N2a cells from nucleic condensation to nucleic fragmentation. Conclusions High-content screening combined with live cell real-time imaging technology is a powerful tool for tracking the morphological variation of apoptoticN2a cells induced by glutamate in vitro, which can also be applied for analyzing the pro or anti- apoptotic function of drugs via tracking the morphological changes of the apoptotic cells.

关 键 词：谷氨酸 神经瘤母细胞N2a 高内涵筛选 变异系数 

分 类 号：Q335[生物学—遗传学] Q26