Rapid 3-D Nano-Printing to Create Multi-Thousand-Channel Microelectrode Arrays

摘要 最近的技术进步使得能够对内部多达几百个神经元的活动进行采样 大脑的一个有限区域。这些创新揭示了理解神经相互作用的重要性 超越局部，微观尺度-并强调目前无法记录足够的组织体积， 理解中尺度和宏观尺度的计算。我们建议克服这一障碍， 通过使用我们最近建立的3D纳米颗粒打印， 法建议的三维阵列将拥有一个数量级以上的记录 现场（5000柄/平方厘米，深度达4毫米），生产成本仅为目前 技术.我们提出的设计克服了该领域的采样和结构的现有限制。 此外，我们的技术有望在建立中观和宏观经济方面带来重要的新见解。 在组织的3D体积内缩放电路动态。 在目标1中，我们将构建第一个大规模微电极阵列（MMEA），能够从5000 通道在1 cm 2和高达4 mm深的组织体积内（柄间距高达140 um）。的 所得到的柄将具有跨越500至4000 μ m的可变长度。这些尺寸使 同时记录小鼠皮质和皮质下结构，具有最小的组织损伤。探针 我们制造的刀柄具有很强的弹性--即使在大位移下也不会断裂 从而在插入过程中容许明显的未对准。尖端的问题也需要巨大的 技术灵活性。我们的生产方法支持不同设计的快速生产 使用基本的AutoCAD软件。此功能可按需进行研究特定的新原型设计， 几个小时内完成电极配置。在目标2中，我们将验证和改进体内探针性能。的 该产品的长期目标是生产一种必不可少的，但价格低廉的神经生理学工具。我们将 建立探针的功能，并最大限度地提高从单个神经元获得的信号的质量， 在小鼠体内。作为第一步，我们将监测头部固定的动物在跑步机上行走时的神经活动。 通过与已经获得的硅探针记录进行比较，这一步骤将验证信号的质量 从不同的大脑区域的不同深度。接下来，我们将光遗传学地唤起神经活动， 神经亚群直接的躯体刺激将提供一个因果关系的反应性措施， 探头，测定其光电灵敏度。

Abstract Recent advances in technology have enabled the sampling of activity from up to a few hundred neurons within a limited area of the brain. These innovations reveal the importance of understanding neural interactions beyond the local, micro-scale – and highlight the current inability to record from sufficient volumes of tissue to understand the meso- and macro-scale computations. We propose to overcome this hurdle, increasing the planar reach and spatial densities of recording sites by using our recently established 3D nanoparticle printing method. The proposed three-dimensional arrays will possess an order of magnitude more recording sites (5000 shanks/cm2 at depths up to 4 mm) at a fraction of the production cost of current technologies. Our proposed design overcomes the field's current limitations of both sampling and structure. Moreover, our technology promises to lead to significant new insights in establishing the meso- and macro- scale circuit dynamics within 3D volumes of tissue. In Aim 1, we will construct the first Massive MicroElectrode Array (MMEA), capable of recording from 5000 channels within a 1 cm2 and up to 4 mm deep volume of tissue (with up to 140 um inter-shank spacing). The resulting shanks will be of variable lengths, spanning 500 to 4000um. These dimensions enable the simultaneous recording of mouse cortical and subcortical structures with minimal tissue damage. The probe shanks we have created are remarkably resilient – bending, but not breaking even under a large displacement and thus tolerating significant misalignment during insertion. Cutting-edge questions also require immense technical flexibility in implementation. Our production method supports the rapid production of different designs using basic AutoCAD software. This feature enables the on-demand, study-specific prototyping of new electrode configurations in a few hours. In Aim 2, we will validate and refine in vivo probe performance. The long-term goal of this product is to produce an essential, yet inexpensive neurophysiology tool. We will establish the probe's functionality and maximize the quality of the signals obtained from individual neurons in vivo in the mouse. As a first step, we will monitor neural activity while a head fixed animal walks on a treadmill. Through comparison with silicon probe recordings already obtained, this step will validate the quality of signal from different depths within individual brain areas. Next, we will optogenetically evoke neural activity from neural sub-populations. Direct somatic stimulation will provide a causal measure of the responsiveness of the probe and determine its photoelectric sensitivity.