Using electrochemical impedance spectroscopy to study biofilm growth in a 3D-printed flow cell system
Using electrochemical impedance spectroscopy to study biofilm growth in a 3D-printed flow cell system
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DOI：
10.1016/j.biosx.2023.100326
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发表时间：
2023-03
影响因子：
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通讯作者：
M. McGlennen;Markus Dieser;C. Foreman;S. Warnat
中科院分区：
文献类型：
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作者：
M. McGlennen;Markus Dieser;C. Foreman;S. Warnat
生物膜污染是一个广泛存在的问题，当生物附着在有水的表面时，它可能发生在任何地方。在工业环境中，生物膜的形成会导致部件失效、材料降解和生物污染或腐败，这些都会带来巨大的经济成本。微制造电化学阻抗谱（EIS）传感器已经成为监测生物膜的一种很有前途的工具，因为EIS传感器可以实时捕获生物膜生长的信息；然而，传感器受到漂移的影响，并且该技术缺乏动态生物膜过程的时间解释。在这项工作中，采用导电聚合物层修饰具有金微交错电极（μIDEs）的微制造传感器，与未修饰的传感器相比，显著降低了EIS测量的可变性，并实现了高度稳定的时间分辨EIS测量。利用一种新型3d打印流细胞系统，利用高分辨率共聚焦激光扫描显微镜（CLSM）对铜绿假单胞菌的生物膜进行了EIS表征，导致EIS数据的明显变化与一致的生物膜生长相对应。我们已经证明，EIS微传感器可以检测生物膜的四个阶段：(i)初始生物膜附着在传感器基质上，（ii）以稀疏生物膜覆盖为特征的早期不可逆生物膜增殖，（iii）以均匀生物膜覆盖为特征的成熟生物膜检测，以及（iv）由于生物膜的剥离和再生而引起的变化。
Biofilm contamination is a widespread issue that can occur anywhere when organisms attach to surfaces in the presence of water. In industrial environments, formation of biofilms can lead to component failure, material degradation, and biofouling or spoilage, which collectively come with significant economic costs. Microfabricated electrochemical impedance spectroscopy (EIS) sensors have emerged as a promising tool for monitoring biofilm as EIS sensors capture information about biofilm growth autonomously in real-time; however, sensors suffer from drift, and the technique lacks temporal interpretation of dynamic biofilm processes. In this work, microfabricated sensors featuring gold micro-interdigitated electrodes (μIDEs) were modified with an electrically conductive polymer layer resulting in EIS measurement variability that was significantly reduced compared to unmodified sensors, and enabled highly stable, time-resolved EIS measurements. EIS characterization ofPseudomonas aeruginosabiofilm in parallel with high-resolution confocal laser scanning microscopy (CLSM) was performed using a novel 3D-printed flow cell system, resulting in distinct changes to EIS data corresponding with consistent biofilm growth. We have shown that EIS microsensors can detect four stages of biofilm: (i) initial biofilm attachment to the sensor substrate, (ii) early-stage irreversible biofilm proliferation characterized by sparse biofilm coverage, (iii) mature biofilm detection characterized by uniform biofilm coverage, and (iv) changes due to detachment and regrowth of biofilm.