Objective: The purpose of this paper is to demonstrate the use of 2-D impedance spectroscopy to identify areas of biofilm growth on a CMOS biosensor microelectrode-array. Methods: This paper presents the design and use of a novel multichannel impedance spectroscopy instrument to allow 2-D spatial and temporal evaluation of biofilm growth. The custom-designed circuits can provide a wide range of frequencies (1 Hz–100 kHz) to allow customization of impedance measurements, as the frequency of interest varies based on the type and state of biofilm under measurement. The device is capable of taking measurements as fast as once per second on the entire set of impedance sensors, allowing real-time observation. It also supports adjustable stimulus voltages. The distance between neighboring sensors is 220 micrometers which provides reasonable spatial resolution for biofilm study. Results: Biofilm was grown on the surface of the chip, occupancy was measured using the new tool, and the results were validated optically using fluorescent staining. The results show that the developed tool can be used to determine the bacterial biofilm presence at a given location. Conclusion: This paper confirms that 2-D impedance spectroscopy can be used to measure biofilm occupancy. The new tool developed to perform the measurements was able to display real-time results, and determine biofilm coverage of the array electrodes. Significance: The system presented in this report is the first fully integrated 2-D EIS measurement system with full software support for capturing biofilm growth dynamics in real-time. Due to its ability to nondestructively monitor biofilms over time, 2-D impedance spectroscopy using a microelectrode-array is a useful tool for studying biofilms.

中文翻译：

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使用 2D 阻抗谱研究细菌生物体的实时时空行为。

客观的： 本文的目的是展示使用二维阻抗谱来识别 CMOS 生物传感器微电极阵列上的生物膜生长区域。 方法：本文介绍了一种新型多通道阻抗光谱仪的设计和使用，以允许对生物膜生长进行二维空间和时间评估。定制设计的电路可以提供宽范围的频率 (1 Hz–100 kHz) 以允许定制阻抗测量，因为感兴趣的频率根据被测生物膜的类型和状态而变化。该设备能够以每秒一次的速度对整套阻抗传感器进行测量，从而实现实时观察。它还支持可调激励电压。相邻传感器之间的距离为 220 微米，这为生物膜研究提供了合理的空间分辨率。结果：生物膜生长在芯片表面，使用新工具测量占用率，并使用荧光染色对结果进行光学验证。结果表明，开发的工具可用于确定给定位置的细菌生物膜存在。结论：本文证实，二维阻抗谱可用于测量生物膜占有率。为执行测量而开发的新工具能够显示实时结果，并确定阵列电极的生物膜覆盖率。意义：本报告中介绍的系统是第一个完全集成的 2-D EIS 测量系统，具有完整的软件支持，可实时捕获生物膜生长动态。由于其能够随时间无损地监测生物膜，使用微电极阵列的二维阻抗谱是研究生物膜的有用工具。