Chronic imaging of neuronal networks in vitro has provided fundamental insights into mechanisms underlying neuronal function. Current labeling and optical imaging methods, however, cannot be used for continuous and long-term recordings of the dynamics and evolution of neuronal networks, as fluorescent indicators can cause phototoxicity. Here, we introduce a versatile platform for label-free, comprehensive and detailed electrophysiological live-cell imaging of various neurogenic cells and tissues over extended time scales. We report on a dual-mode high-density microelectrode array, which can simultaneously record in (i) full-frame mode with 19,584 recording sites and (ii) high-signal-to-noise mode with 246 channels. We set out to demonstrate the capabilities of this platform with recordings from primary and iPSC-derived neuronal cultures and tissue preparations over several weeks, providing detailed morpho-electrical phenotypic parameters at subcellular, cellular and network level. Moreover, we develop reliable analysis tools, which drastically increase the throughput to infer axonal morphology and conduction speed.

体外神经元网络的慢性成像提供了对神经元功能潜在机制的基本见解。然而，目前的标记和光学成像方法不能用于神经元网络动态和演化的连续和长期记录，因为荧光指示剂会引起光毒性。在这里，我们介绍了一个多功能平台，用于在延长的时间范围内对各种神经源性细胞和组织进行无标记、全面和详细的电生理活细胞成像。我们报告了一种双模式高密度微电极阵列，它可以在 (i) 具有 19,584 个记录站点的全帧模式和 (ii) 具有 246 个通道的高信噪比模式下同时进行记录。我们着手展示该平台的功能，在数周内记录了原代和 iPSC 衍生的神经元培养物和组织制备物，提供了亚细胞、细胞和网络级别的详细形态电表型参数。此外，我们开发了可靠的分析工具，大大增加了推断轴突形态和传导速度的吞吐量。