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 H2O2, and a higher sensitivity of 207 units per mg/l of zymosan. Thereafter, a factor of 84 molecules of H2O2 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.

中文翻译：

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一种电化学发光传感矩阵，用于实时探测细胞输出的活性氧种类。

本文中，开发了一种新型的基于细胞的电化学发光（ECL）传感矩阵，用于探测从小鼠巨噬细胞产生的活性氧（ROSs）。通过水解3-氨基丙基三甲氧基硅烷（APTMS）作为传感器基板，在铟锡氧化物（ITO）玻璃表面上装饰平均直径为16 nm的均匀大小的Au纳米粒子（AuNPs）。然后，表面覆盖有聚-1-赖氨酸薄膜，在其中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貌进行表征，并通过电化学阻抗谱研究其电化学性能。从AuNPs / APTMS / ITO底物观察到线性响应，灵敏度为0。每毫克/升H2O2 465单位，灵敏度更高，每毫克/升酵母聚糖207单位。此后，估计由单一糖原产生的84个分子的H2O2分子。结果表明，这种基于细胞的传感器的ECL响应在酵母聚糖的刺激下与细胞氧代谢期间产生的ROS定量相关。这项工作表明，所制备的传感矩阵可有效监测活细胞的氧代谢，并可应用于生物学和临床领域，以提供有关细胞正常或异常功能的重要信息。结果表明，这种基于细胞的传感器的ECL响应在酵母聚糖的刺激下与细胞氧代谢期间产生的ROS定量相关。这项工作表明，所制备的传感矩阵可有效监测活细胞的氧代谢，并可应用于生物学和临床领域，以提供有关细胞正常或异常功能的重要信息。结果表明，这种基于细胞的传感器的ECL响应在酵母聚糖的刺激下与细胞氧代谢期间产生的ROS定量相关。这项工作表明，所制备的传感矩阵可有效监测活细胞的氧代谢，并可应用于生物学和临床领域，以提供有关细胞正常或异常功能的重要信息。