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用于实时探测细胞输出活性氧的电化学发光传感矩阵。

An electrochemiluminescent sensing matrix for real-time probing of cell-output reactive oxygen species.

作者信息

Fang Chen, Du Dan, Li Dongning, Huang Jie, Tu Yifeng

机构信息

College of Chemistry, Chemical Engineering and Material Science, Dushu Lake Campus, Soochow University, Industrial Park, Suzhou 215123, People's Republic of China.

School of Biology & Basic Medical Science, Dushu Lake Campus, Soochow University, Industrial Park, Suzhou 215123, People's Republic of China.

出版信息

Biomicrofluidics. 2019 Aug 29;13(4):044115. doi: 10.1063/1.5111379. eCollection 2019 Jul.

Abstract

Herein, a novel cell-based electrochemiluminescent (ECL) sensing matrix was developed for probing reactive oxygen species (ROSs) produced from mouse macrophage cells. Uniformly sized Au nanoparticles (AuNPs) with an average diameter of 16 nm were decorated on the surface of indium tin oxide (ITO) glass through the connection of hydrolyzed 3-aminopropyl trimethoxysilane (APTMS) serving as a sensor substrate. Then, the surface was covered with a poly-l-lysine thin film, where mouse macrophage cells were successfully cultured. The morphology of the electrodes obtained was characterized by scanning electron microscopy and atomic force microscopy, and their electrochemical properties were investigated by electrochemical impedance spectroscopy. A linear response was observed from the AuNPs/APTMS/ITO substrate with a sensitivity of 0.465 units per mg/l of HO, and a higher sensitivity of 207 units per mg/l of zymosan. Thereafter, a factor of 84 molecules of HO produced by a single glycogen was estimated. The results demonstrated that the ECL response of this cell-based sensor quantitatively correlated with yielded ROSs during cell oxygen metabolism under the stimulation of zymosan. This work suggests that the prepared sensing matrix is efficient for monitoring the oxygen metabolism of living cells and can be applied in biological and clinical fields to provide significant information on the regular or abnormal function of cells.

摘要

在此,开发了一种新型的基于细胞的电化学发光(ECL)传感基质,用于探测小鼠巨噬细胞产生的活性氧(ROS)。通过连接水解的3-氨丙基三甲氧基硅烷(APTMS)作为传感器底物,将平均直径为16nm的均匀尺寸金纳米颗粒(AuNP)装饰在氧化铟锡(ITO)玻璃表面。然后,表面覆盖有聚-L-赖氨酸薄膜,在该薄膜上成功培养了小鼠巨噬细胞。通过扫描电子显微镜和原子力显微镜对所得电极的形态进行了表征,并通过电化学阻抗谱研究了它们的电化学性质。在AuNP/APTMS/ITO底物上观察到线性响应,对HO的灵敏度为每mg/l 0.465单位,对酵母聚糖的灵敏度更高,为每mg/l 207单位。此后,估计单个糖原产生的HO分子数为84个。结果表明,这种基于细胞的传感器的ECL响应与酵母聚糖刺激下细胞氧代谢过程中产生的ROS定量相关。这项工作表明,所制备的传感基质对于监测活细胞的氧代谢是有效的,可应用于生物和临床领域,以提供有关细胞正常或异常功能的重要信息。

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