高等学校化学学报 ›› 2005, Vol. 26 ›› Issue (2): 247.

• 研究简报 • 上一篇    下一篇

微流控芯片NDA在线衍生测定单细胞中谷胱甘肽

凌云扬, 殷学锋, 方肇伦   

  1. 浙江大学化学系微分析系统研究所, 杭州 310027
  • 收稿日期:2004-02-12 出版日期:2005-02-10 发布日期:2005-02-10
  • 通讯作者: 殷学锋(1945年出生),男,教授,博士生导师,从事微流控分析芯片研究.E-mail:yinxf@sun.zju.edu.cn E-mail:yinxf@sun.zju.edu.cn
  • 基金资助:

    国家自然科学基金重大项目(批准号:20299030)资助

On-chip Labeling with NDA for Determining Glutathione in Single Cell Using Microfluidic Chip Electrophoresis

LING Yun-Yang, YIN Xue-Feng, FANG Zhao-Lun   

  1. Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou 310027, China
  • Received:2004-02-12 Online:2005-02-10 Published:2005-02-10

关键词: 微流控芯片, 单细胞, 在线衍生, 谷胱甘肽

Abstract: A simple method for the detection of glutathione(GSH) in single human erythrocyte was developed using on-chip mode dynamic 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 laser induced fluorescence(LIF) detection. By using a combination of hydrostatic pressure and low electric field, single cell sampling speed and long-term stability were improved. 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 the microchip separation channel during electrophoresis. The average separation efficiency for GSH was 2.4×106 plates/m, which was not a significant difference from off-chip derivatization. The GSH migration time on the on-chip mode and off-chip method were 116 and 110 s, respectively. The present method can be used to minimize the dilution of the contents of single cell during the derivatization to maintain favorable kinetics for the labeling reaction, 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.

Key words: Microfluidic chip, Single cell, On-chip derivatization, Glutathione

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