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

我们中枢神经系统的神经功能是由庞大的神经网络中不同类型的神经元之间复杂的相互作用产生的。为了了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。这可以通过成像（可视化各种细胞类型中表达的荧光标记）或电极阵列（多电极阵列；MEAs）同时记录数百到数千个神经元的电活动来实现（例如在脑切片中），这些神经元在功能上相互连接（或结合这两种方法）。当使用大脑切片时，由于切片过程破坏了细胞过程的完整性，表面细胞受到损伤。因此，这些记录通常是通过穿透电极或光学记录在组织深处完成的。传统的mea由平面电极组成，只能从组织表面进行记录。它们用于记录分离培养的细胞，生长在电极上，或从分离的视网膜上，输出细胞形成单一的浅层。但这些平面mea不适于从脑切片记录。目前市场上大多数带有穿透电极的MEA系统对于精致的孤立神经组织来说都太大了，而且它们被限制在不超过100个电极，这不足以分析我们在我们提议的相关项目中研究的网络动力学。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4096个密集的电极组成。允许记录间隔的距离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，因此可以在细胞不受损、神经网络完整的情况下记录深度神经活动。我们将把这个尖端的新记录系统与荧光显微镜结合起来，这将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并发成像。这个新颖的平台将允许我们在宏观尺度上研究健康和疾病中的神经功能，使用大脑切片，视网膜（来自啮齿动物和人类组织）和人类干细胞衍生的人造器官（视网膜，内耳）。我们期待这个新设施将在纽卡斯尔吸引令人兴奋的新合作和技术发展。

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.