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首页> 外文期刊>Biosensors & Bioelectronics: The International Journal for the Professional Involved with Research, Technology and Applications of Biosensers and Related Devices >Superoxide microsensor integrated into a Sensing Cell Culture Flask microsystem using direct oxidation for cell culture application
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Superoxide microsensor integrated into a Sensing Cell Culture Flask microsystem using direct oxidation for cell culture application

机译：使用直接氧化技术将超氧化物传感器集成到传感细胞培养瓶微系统中，用于细胞培养

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A new electrochemical sensor system for reliable and continuous detection of superoxide radical release from cell culture was developed utilizing direct oxidation of superoxide on polymer covered gold microelectrodes. Direct superoxide oxidation was demonstrated to provide robust measurement principle for sensitive and selective reactive oxygen species (ROS) quantification without the need for biocomponent supported conversion. Sensor performance was investigated by using artificial enzymatic superoxide production revealing a sensitivity of 2235 A M-1 m(-2). An electrode protection layer with molecular weight cut-off property from adsorbed linear branched polyethylenimine was successfully introduced for long term and selectivity improvement. Thin-film based sensor chip fabrication with implemented three-electrode setup and full integration into the technological platform Sensing Cell Culture Flask was described. Cell culturing directly on-chip and free radical release by phorbol-12-myristate-13-acetate (PMA) stimulation was demonstrated using T-47D human breast cancer carcinoma cell model. Transient extracellular superoxide production upon stimulation was successfully observed from amperometric monitoring. Signal inhibition from scavenging of extracellular superoxide by specific superoxide dismutase (SOD) showed the applicability for selective in vitro ROS determination. The results confirm the possibility of direct superoxide oxidation, with exclusion of the main interfering substances uric acid and hydrogen peroxide. This offers new insights into the development of reliable and robust ROS sensors. (C) 2014 Elsevier B.V. All rights reserved.
机译：利用在聚合物覆盖的金微电极上直接氧化超氧化物，开发了一种新的电化学传感器系统，用于可靠连续地检测细胞培养物中的超氧化物自由基释放。直接超氧化物氧化被证明为灵敏和选择性的活性氧（ROS）定量提供了可靠的测量原理，而无需生物组分支持的转化。通过使用人工酶超氧化物生产调查了传感器性能，揭示了2235 A M-1 m（-2）的灵敏度。成功地从吸附的线性支链聚乙烯亚胺中引入了一种具有分子量截断特性的电极保护层，以长期有效地提高选择性。描述了基于薄膜的传感器芯片的制造，该传感器芯片具有实现的三电极设置并完全集成到了传感细胞培养瓶技术平台中。使用T-47D人乳腺癌癌细胞模型，证实了细胞在片上直接培养以及佛波12-肉豆蔻酸13-乙酸酯（PMA）刺激下的自由基释放。从安培监测成功观察到刺激后瞬时细胞外超氧化物的产生。通过特异性超氧化物歧化酶（SOD）清除细胞外超氧化物的信号抑制显示了选择性体外ROS测定的适用性。结果证实了直接超氧化物氧化的可能性，排除了主要干扰物质尿酸和过氧化氢。这为开发可靠而强大的ROS传感器提供了新见解。 （C）2014 Elsevier B.V.保留所有权利。

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