AI 中文总结
该研究开发了集成纳米多孔膜的高内涵TEER装置，结合Conv1d和KAN模型，可原位监测片上屏障动态，实现细胞层形成阶段及屏障弱化的高置信度识别，无需显微和终点染色。
AI 中文摘要
传统跨上皮电阻（TEER）技术仅能对细胞层状态进行低内涵分析，对于片上屏障系统而言，需在培养箱外重复进行形态学和细胞间连接的显微评估。本研究提出一种新型高内涵TEER装置，采用新型纳米多孔膜，可实现细胞-基底阻抗传感（ECIS）的连续电学测量。该超薄膜厚度为700 nm，由超低应力SixNy构成，单片集成于晶圆级制造、带玻璃盖密封的芯片中。共面ECIS电极连接定制电子设备，用于记录正弦激励下的阻抗。接种人脐静脉内皮细胞（HUVEC）后，将连续记录的阻抗谱与明场、荧光显微镜结果对比，可识别单层形成的不同阶段。使用少量奈奎斯特图训练的一维卷积神经网络（Conv1d）和柯尔莫哥洛夫-阿诺德网络（KAN），能以95%的置信度识别出四个阶段：（I）黏附、（II）铺展、（III）汇合、（IV）紧密连接形成的屏障成熟。作为进一步的概念验证，该方法可识别调节剂PN159和BAC诱导的可逆及不可逆屏障弱化。本研究证明，该技术可即时、自动化、非侵入式地检测片上屏障系统内的体外屏障动态，无需显微和终点染色。我们预计该ECIS技术将在器官芯片系统中广泛应用，用于原位监测组织屏障的生理或病理状态。
英文摘要
Conventional transepithelial electrical resistance (TEER) technique provides only a low-content analysis of cell-layer conditions, necessitating repeated microscopic assessments of morphology and cell-cell contacts outside the incubator for barrier-on-chip systems. This work presents a novel high-content TEER device in the form of a novel nanoporous membrane that facilitates continuous electrical measurement of cell-substrate impedance sensing (ECIS). The ultrathin (700 nm) membrane, composed of ultra-low-stress SixNy, is monolithically integrated into wafer-level fabricated chips sealed with glass lids. Coplanar ECIS electrodes were connected to custom electronics to record impedance under sinusoidal excitation. Human umbilical vein endothelial cells (HUVECs) were seeded and continuously recorded impedance spectra were compared with bright-field and fluorescence microscopy, revealing distinct phases of monolayer formation. With one-dimensional convolutional neural network (Conv1d) and Kolmogorov-Arnold Network (KAN) trained with a small amount of Nyquist-diagrams, phases of (I) adherence, (II) outspreading, (III) confluence and (IV) barrier maturity with tight junction formation could be recognized with 95% confidence. As further proof of concept, reversible and irreversible barrier weakening using modulators PN159 and BAC was identified in this way. Our studies have demonstrated that an immediate and automatable non-invasive detection of in-vitro barrier dynamics within barrier-on-chip systems, eliminating the need for microscopy and endpoint staining. We expect this ECIS technique will find broad applications in organ-on-chip systems for in situ monitoring physiological or pathological states of tissue barrier.
