Simultaneous quantification of multiple endogenous biothiols in single living cells by plasmonic Raman probes,Chemical Science

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Simultaneous quantification of multiple endogenous biothiols in single living cells by plasmonic Raman probes
Chemical Science ( IF 7.6 ) Pub Date : 2017-08-29 00:00:00 , DOI: 10.1039/c7sc03218h
Shan-Shan Li _{1,

2,

3,

4,

5} , Qi-Yuan Guan _{1,

2,

3,

4,

5} , Mengmeng Zheng _{1,

2,

3,

4,

5} , Yu-Qi Wang _{1,

2,

3,

4,

5} , Deju Ye _{1,

2,

3,

4,

5} , Bin Kang _{1,

2,

3,

4,

5} , Jing-Juan Xu _{1,

2,

3,

4,

5} , Hong-Yuan Chen _{1,

2,

3,

4,

5}

Affiliation

Intracellular biothiols mediate many important physiological and pathological processes. Due to their low content and competing thiol-reactivity, it is still an unmet challenge to quantify them within a complicated intracellular environment. Herein, we demonstrated a strategy to discriminate three biothiols, i.e. cysteine (Cys), homo-cysteine (Hcy) and glutathione (GSH), and quantify their concentrations within single living cells, using one platform of Raman probe. By monitoring the reaction kinetics of biothiols with Raman probes and discriminating their products with a quantitative principal component analysis (qPCA) method, these three biothiols could be simultaneously quantified in both cell lysis and single living cells. The concentrations of Cys, Hcy and GSH in single Hela cells were 158 ± 19 μM, 546 ± 67 μM and 5.07 ± 0.62 mM, respectively, which gives the precise concentrations of these three biothiols at a single cell level for the first time. This method provides a general strategy for discriminating each component from a mixed system and has potential for quantifying any biomolecules within an in vitro or in vivo biological environment.

中文翻译：

等离子体拉曼探针同时定量单个活细胞中多种内源性生物硫醇

细胞内生物硫醇介导许多重要的生理和病理过程。由于它们的含量低和竞争性的硫醇反应性，在复杂的细胞内环境中对其进行定量仍然是一项尚未解决的挑战。在这里，我们展示了一种区分三种生物硫醇的策略，即半胱氨酸（Cys），高半胱氨酸（Hcy）和谷胱甘肽（GSH），并使用一个拉曼探针平台量化它们在单个活细胞中的浓度。通过用拉曼探针监测生物硫醇的反应动力学并通过定量主成分分析（qPCA）方法区分其产物，可以同时定量分析细胞裂解液和单个活细胞中的这三种生物硫醇。单个Hela细胞中Cys，Hcy和GSH的浓度分别为158±19μM，546±67μM和5.07±0.62 mM，这是第一次在单个细胞水平上精确给出了这三种生物硫醇的浓度。该方法提供了一种从混合系统中区分每种成分的一般策略，并且具有定量体外或体外生物分子的潜力。体内生物环境。

更新日期：2017-09-22

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