Malerba Mario, Amin Hayder, Angotzi Gian N, Maccione Alessandro, Berdondini Luca
Fondazione Istituto Italiano di Tecnologia (IIT), Neuroscience and Brain Technologies, Genoa, Italy.
Methods Mol Biol. 2018;1771:147-157. doi: 10.1007/978-1-4939-7792-5_12.
Substrate-integrated multielectrode arrays (MEAs) enable multisite, long-term, and label-free sensing and actuation of neuronal electrical signals in reduced cell culture models for network electrophysiology. Conventional, thin-film fabricated passive MEAs typically provide a few tens of electrode sites. New generations of active CMOS-based high-resolution arrays provide the capabilities of simultaneous recordings from thousands of neurons over fields of view of several square millimeters, yet allowing extracellular electrical imaging to be achieved down to the subcellular scale. In turn, such advancement in chip-based electrical readouts can significantly complement recently developed biotechnological and bimolecular techniques for neurobiology applications. Here, we describe (1) a simple method to fabricate passive MEAs and (2) protocols for preparing and growing primary rat hippocampal neuronal cultures and human iPS-derived neurons on MEAs. The aim is to provide reliable protocols for initiating the reader to this technology and for stimulating their further development and experimental use in neurobiology.
衬底集成多电极阵列(MEA)能够在用于网络电生理学的简化细胞培养模型中对神经元电信号进行多部位、长期且无标记的传感与驱动。传统的薄膜制造无源MEA通常提供几十个电极位点。新一代基于有源CMOS的高分辨率阵列具备在数平方毫米视野范围内同时记录数千个神经元的能力,还能实现亚细胞尺度下的细胞外电成像。反过来,基于芯片的电读出技术的这种进步可以显著补充最近为神经生物学应用开发的生物技术和双分子技术。在此,我们描述(1)一种制造无源MEA的简单方法,以及(2)在MEA上制备和培养原代大鼠海马神经元培养物和人诱导多能干细胞衍生神经元的方案。目的是提供可靠的方案,使读者初步了解这项技术,并促进其在神经生物学中的进一步发展和实验应用。