微流控芯片NDA在线衍生测定单细胞中谷胱甘肽

单细胞分析对研究细胞内信号传递和重大疾病的早期诊断等具有重要意义，荧光标记是检测细胞内物质的常用技术，为防止衍生时的过度稀释，大多采用柱前细胞内衍生法，衍生后再用微流控芯片分析，此法操作复杂，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。

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

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.