State Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources, College of Chemistry and pharmacy, Guangxi Normal University, Guilin 541004, China; Guilin Normal College, Guilin 541001, China.
State Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources, College of Chemistry and pharmacy, Guangxi Normal University, Guilin 541004, China.
Talanta. 2018 Mar 1;179:466-471. doi: 10.1016/j.talanta.2017.11.044. Epub 2017 Nov 22.
Glutathione (GSH) is a major endogenous antioxidant that has a central role in cellular defense against toxins and free radicals. Rapid and accurate detection of GSH content in single cells is important to the early diagnosis of disease and biomedical research. In this work, a novel method based on microchip electrophoresis chemiluminescence (MCE-CL) detection was developed for the quantification of glutathione (GSH) in single cells from rat liver. The detection of GSH is based on the strong sensitization of mercapto compound to luminol-HOCL system. The injection, localization, and membrane dissolution of single cell were simply and rapidly carried out on the microchip by direct electric field force, which did not require any additional membrane dissolution reagent. Under optimized experimental conditions, single cell assay was achieved within 2min. The peak area of the GSH was taken as quantification of GSH, and a good linear relationship of GSH concentration to peak area in the range of 3.0 × 10M to 6.0 × 10M was obtained. The detection limit for GSH is 9.6 × 10M, calculated by S/N = 3. The measured GSH content in single cells from rat liver (n = 10) ranged from 7.8fmol to 13.fmol with a mean value of 10.8fmol.
谷胱甘肽（GSH）是一种主要的内源性抗氧化剂，在细胞防御毒素和自由基方面起着核心作用。快速准确地检测单个细胞中的 GSH 含量对于疾病的早期诊断和生物医学研究非常重要。在这项工作中，开发了一种基于微芯片电泳化学发光（MCE-CL）检测的新方法，用于定量测定大鼠肝单个细胞中的谷胱甘肽（GSH）。GSH 的检测基于巯基化合物对鲁米诺-HOCL 体系的强烈敏化作用。通过直接电场力，在微芯片上简单快速地进行单细胞的注入、定位和膜溶解，而不需要任何额外的膜溶解试剂。在优化的实验条件下，在 2 分钟内完成了单细胞测定。将 GSH 的峰面积作为 GSH 的定量，在 3.0×10^-6M 至 6.0×10^-6M 的范围内，GSH 浓度与峰面积之间存在良好的线性关系。GSH 的检测限为 9.6×10^-6M，按 S/N = 3 计算。从大鼠肝的单个细胞中（n = 10）测得的 GSH 含量范围为 7.8fmol 至 13.fmol，平均值为 10.8fmol。
