在线衍生微流控芯片电泳和激光诱导荧光法测定单细胞中谷胱甘肽

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^1~4].在测定细胞内容物时,大多采用柱前细胞内衍生法^1,2,4],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.

On-Chip Derivatization for Determining Glutathione in Single Cell Using Microchip Electrophoresis and Laser Induced Fluorescence Detection

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 2.4×10⁶ plates/m) which was not significantly different from off-chip derivatization.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 derivatization,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.