Multidimensional large-scale, high-density in vitro recording facility for the investigation of neural systems function

我们中枢神经系统中的神经功能源于巨大神经网络中不同神经元类型之间的复杂相互作用。为了理解这些网络是如何运作的，同时记录大量神经元群是必不可少的。这可以使用成像（以可视化在各种细胞类型中表达的荧光标记物）或电极阵列（多电极阵列; MEA）来离体（例如在脑切片中）实现，电极阵列同时记录数百至数千个功能上相互连接的神经元的电活动（或结合两种方法）。当使用脑切片时，表面细胞会受到损伤，因为切片过程破坏了细胞过程的完整性。由于这个原因，这些记录通常是在组织的深处用穿透电极或光学记录完成的。传统的MEA由只能从组织表面记录的平面电极组成。它们可用于记录在电极上生长的分离培养物中的细胞，或来自分离的视网膜，其中输出细胞形成单个表面层。但是这些平面MEA不适合从脑切片记录。目前市场上大多数具有穿透电极的MEA系统对于脆弱的孤立神经组织来说都太大了，并且它们被限制为具有不超过100个电极，这不足以分析我们在我们提出的相关项目中旨在研究的网络动力学。通过这个提议，我们的目标是围绕一个独特的下一代MEA系统建立一个设施，该系统由4,096个密集的电极组成，允许记录间隔与神经网络中相邻神经元之间的距离相似。这些电极由穿透组织的柱子组成，因此可以深度记录神经活动，其中细胞没有受损，神经网络完好无损。我们将联合收割机与荧光显微镜相结合，使我们能够在神经元相互传递信号的同时对各种生理参数进行同步成像。这个新颖的平台将使我们能够使用脑切片、视网膜和神经元切片，在宏观尺度上研究健康和疾病中的神经功能。（从啮齿动物和人体组织）和人类干细胞衍生的人工器官（视网膜，内耳）。我们预计，这个新的设施将吸引令人兴奋的新的合作和技术发展在纽卡斯尔。

Neural function in our central nervous system arises from complex interactions between different neuronal types within vast neural networks. To understand how these networks operate, simultaneous recordings from large neuronal populations are essential. This can be achieved ex vivo (for example in brain slices) using either imaging (to visualise fluorescent markers expressed in various cell types), or arrays of electrodes (multielectrode arrays; MEAs) that record electrical activity simultaneously from hundreds to thousands of neurons that are functionally connected to each other (or combining both approaches). When using brain slices, surface cells are damaged because of the slicing process that disrupts the integrity of their cellular processes. For that reason, these recordings are normally done in the depth of the tissue with penetrating electrodes or with optical recordings.Traditional MEAs consist of planar electrodes that can record only from the surface of the tissue. They are useful to record from cells in dissociated cultures, growing on the electrodes, or from the isolated retina, where the output cells form a single superficial layer. But these planar MEAs are not amenable to record from brain slices. Most MEA systems currently on the market with penetrating electrodes are too large for delicate isolated neural tissue, and they are limited to having no more than 100 electrodes, which is insufficient to analyse the network dynamics that we aim to study in the related projects we propose..With this proposal, we aim to establish a facility around a unique next-generation MEA system consisting of 4,096 densely packed electrodes, allowing recordings spaced at similar distance to that between adjacent neurones in neural networks. These electrodes consist of pillars that penetrate the tissue and can therefore record neural activity in depth, where cells are not damaged and neural networks are intact. We will combine this cutting-edge new recording system with a fluorescent microscope that will allow us to perform concurrent imaging of various physiological parameters while neurons signal to each other.This novel platform will allow us to study neural function in health and disease at macroscopic scale in health and disease using brain slices, retinas (from rodents and human tissue) and human stem-cell-derived artificial organs (retina, inner-ear).We expect that this new facility will attract exciting new collaborations and technology developments in Newcastle.