【作者】 凌云扬； 殷学锋； 方肇伦；

【Author】 LING Yun-Yang, YIN Xue-Feng, FANG Zhao-Lun(Institute of Microanalysis System, Department of Chemistry, Zhejiang University,Hangzhou 310027, China)

【机构】 浙江大学化学系微分析系统研究所；

【摘要】 <正> 微流控分析芯片的网络结构和微米通道尺寸适合于单细胞进样、控制和分离分析.在测定细胞内容物时,大多采用柱前细胞内衍生法,但操作复杂,需多次离心分离,且能透过细胞膜标记胞内组分的荧光试剂较少.本文通过将衍生剂NDA加入电泳缓冲液,在线动态标记了单细胞中的谷胱甘肽,节约了衍生试剂,简化了细胞的处理方法,提高了分析速度.为微流控芯片上单细胞内组分分析提供了一种新的衍生方法.实现了微流控芯片上单细胞进样、贴壁、溶膜、衍生、分离和测定的集成化.每小时可分析20个单细胞.更多还原

【Abstract】 A simple method for the detection of glutathione (GSH) in single human erythrocyte was developed by using on-chip labeling by adding the 2,3-naphthalene-dicarboxaldehyde(NDA) simply in microchip electrophoresis buffer, with functional integration of cell sampling, single cell loading docking , lysing, dynamic labeling, electrophoresis separation and LIF detection. After lysing, the reaction between the released GSH and NDA included in electrophoresis buffer can be completed within a migrating distance of 0. 5 cm in separation channel during electrophoresis. The average separation efficiency for GSH was 2. 4×106plates/m, which was not significantly different from off-chip derivatiza-tion. The GSH migration time on the on-chip mode and off-chip method were 116 s and 110 s, respectively. The present method can minimize the dilution of the contents of single cell during derivatiza-tion, simplify the cell treatment and save the sample and reagent. A half life of 5 days of GSH in human erythrocyte was found by using this method.更多还原