实时成像和流式细胞术检测活细胞中的谷胱甘肽含量。
Glutathione Quantification in Live Cells with Real-Time Imaging and Flow Cytometry.
机构信息
Department of Pharmacology and Chemical Biology, Baylor College of Medicine, Houston, TX 77030, USA.
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USA.
出版信息
STAR Protoc. 2020 Nov 14;1(3):100170. doi: 10.1016/j.xpro.2020.100170. eCollection 2020 Dec 18.
Glutathione (GSH) is a highly dynamic, high abundance molecule regulating redox homeostasis in most mammalian cells. Traditional methods could not achieve quantification of glutathione in live cells with high spatial and temporal resolution. Here, we provide protocols on how to use reversible reaction-based ratiometric fluorescent probes, RealThiol (RT) and its derivatives, to quantify GSH globally or in specific organelles. The protocols are applicable to cultured or harvested cells through confocal imaging and flow cytometry. For complete details on the use and execution of this protocol, please refer to Chen et al. (2017) and Jiang et al. (2015, 2017, 2018a).
谷胱甘肽（GSH）是一种高度动态、高丰度的分子，可调节大多数哺乳动物细胞中的氧化还原稳态。传统方法无法实现对活细胞中谷胱甘肽进行高时空分辨率的定量。在这里，我们提供了使用基于可逆反应的比率荧光探针 RealThiol（RT）及其衍生物来定量测量细胞内或细胞器内 GSH 的总体水平或特定水平的方案。该方案可通过共聚焦成像和流式细胞术应用于培养或收获的细胞。如需详细了解本方案的使用和执行情况，请参考 Chen 等人（2017）和 Jiang 等人（2015、2017、2018a）的研究。
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