Xu Xingyuan, Liu Zhengjie, Liu Jing, Yao Chuanjie, Chen Xi, Huang Xinshuo, Huang Shuang, Shi Peng, Li Mingqiang, Wang Li, Tao Yu, Chen Hui-Jiuan, Xie Xi
State Key Laboratory of Optoelectronic Materials and Technologies, Guangdong Province Key Laboratory of Display Material and Technology, School of Electronics and Information Technology, Sun Yat-Sen University, Guangzhou, 510006, China.
The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510080, China.
Microsyst Nanoeng. 2025 May 13;11(1):85. doi: 10.1038/s41378-025-00887-6.
Microelectrode arrays (MEAs) are essential tools for studying the extracellular electrophysiology of cardiomyocytes in a multi-channel format. However, they typically lack the capability to record intracellular action potentials (APs). Recent studies have relied on costly fabrication of high-resolution microelectrodes combined with electroporation for intracellular recordings, but the impact of microelectrode size on micro-electroporation and the quality of intracellular signal acquisition has yet to be explored. Understanding these effects could facilitate the design of microelectrodes of various sizes to enable lower-cost manufacturing processes. In this study, we investigated the influence of microelectrode size on intracellular AP parameters and recording metrics post-micro-electroporation through simulations and experiments. We fabricated microelectrodes of different sizes using standard photolithography techniques to record cardiomyocyte APs from various culture environments with coupled micro-electroporation. Our findings indicate that larger microelectrodes generally recorded electrophysiological signals with higher amplitude and better signal-to-noise ratios, while smaller electrodes exhibited higher perforation efficiency, AP duration, and single-cell signal ratios. This work demonstrates that the micro-electroporation technique can be applied to larger microelectrodes for intracellular recordings, rather than being limited to high-resolution designs. This approach may provide new opportunities for fabricating microelectrodes using alternative low-cost manufacturing techniques for high-quality intracellular AP recordings.
微电极阵列（MEAs）是以多通道形式研究心肌细胞细胞外电生理学的重要工具。然而，它们通常缺乏记录细胞内动作电位（APs）的能力。最近的研究依赖于高分辨率微电极的昂贵制造，并结合电穿孔进行细胞内记录，但微电极尺寸对微电穿孔和细胞内信号采集质量的影响尚未得到探索。了解这些影响有助于设计各种尺寸的微电极，以实现低成本的制造工艺。在本研究中，我们通过模拟和实验研究了微电极尺寸对微电穿孔后细胞内AP参数和记录指标的影响。我们使用标准光刻技术制造了不同尺寸的微电极，通过耦合微电穿孔从各种培养环境中记录心肌细胞的APs。我们的研究结果表明，较大的微电极通常记录到的电生理信号幅度更高、信噪比更好，而较小的电极则表现出更高的穿孔效率、AP持续时间和单细胞信号比率。这项工作表明，微电穿孔技术可应用于较大的微电极进行细胞内记录，而不仅限于高分辨率设计。这种方法可能为使用替代低成本制造技术制造微电极以进行高质量细胞内AP记录提供新的机会。
