Development of 3D-FAST Optical Interface for Rapid Volumetric Neural Sensing and Modulation

项目摘要 为了加深我们对神经回路功能的理解，需要新的工具来 从共同神经中涉及的大量神经元收集同步测量数据 计算并提供精确的功能调节。清醒动物表达的光学成像 钙或电压指示器提供来自个人的实时功能和空间信息 局部神经回路中的神经元。当前成像技术的局限性包括小范围的 包括单个大脑区域的视图，以及头部固定的要求，这防止了 自然主义的行为。此外，大多数光学成像系统不允许同时高电平 结合空间局域光遗传调制的分辨率函数成像。 为了迎接这一挑战，我们建议开发一种能够快速、实时地- 时间容积神经记录和精确的光学刺激。通过将微型阵列配对 微图案化LED发射器具有电润湿透镜技术的轴向聚焦能力，我们 将实现对数千个神经元的双重记录和刺激。通过对小说的利用 3D打印脚手架，我们将能够创建模块化、可扩展、可定制的镜头阵列， 允许记录大规模的双向神经接口，用于神经的闭环调制 电路。 我们将创建两个版本的3D-FAST设备。3D-FAST-GECI将提供快速、体积 GCaMP成像的30赫兹成像，以及3D-FAST-GEVI，它将提供1 kHz的帧速率 单平面与较慢的高分辨率体积扫描相结合。最初，这些设备将在 麻醉，然后清醒，自由移动的小鼠，用于使用GCaMP电压指示器进行实验 ASAP和Voltron，最后结合本地化光遗传学进行闭环反馈。 总而言之，这些实验将展示3D-FAST技术的独特能力。迅速， 改进的电压指示器的成像将与空间受限的光传输相结合，以 首次在自由活动的小鼠中进行光发生神经调制。光学成像特性 将与地面真相双光子显微镜进行比较，以及功能结果 神经调节将通过行为分析进行剖析。3D-FAST工具将带来新的 测量和调节自由活动动物的大量神经元的能力，以更好地 理解行为背后的神经计算。此外，这项工作将为 为未来开发全植入式光学记录和调制单元奠定了基础 自由移动、不受束缚的自然主义行为实验。

Project Summary To further our understanding of the function of neural circuits, there is a need for new tools that can collect simultaneous measurements from large populations of neurons involved in a common neural computation and provide precise functional modulation. Optical imaging in awake animals expressing calcium or voltage indicators provides real-time functional and spatial information from individual neurons within local neural circuits. The limitations of current imaging technology include small fields of view encompassing single brain regions, and the requirement for head fixation, which prevents naturalistic behavior. In addition, most optical imaging systems do not allow for simultaneous high- resolution functional imaging in combination with spatially-localized optogenetic modulation. To meet this challenge, we propose to develop an optical device (`3D-FAST') that allows for rapid, real- time volumetric neural recording and precise optical stimulation. By pairing miniature arrays of micropatterned LED emitters with the axial focusing capabilities of electrowetting lens technologies, we will achieve duplex recording and stimulation of many thousands of neurons. Through utilization of novel 3D-printed scaffolding, we will be able to create modular, expandable, customizable lens arrays that allow for recording of large-scale bi-directional neural interfaces for closed-loop modulation of neural circuits. We will create two versions of the 3D-FAST device. 3D-FAST-GECI that will provide fast, volumetric imaging at 30 Hz for GCaMP imaging, and 3D-FAST-GEVI that will provide frame rates of 1 kHz at a single plane combined with a slower, high resolution volume scan. Initially, the devices will be tested in the anesthetized, and then awake, freely moving mouse for experiments using GCaMP, voltage indicators ASAP and Voltron, and finally combining with localized optogenetics for closed loop feedback. In sum, these experiments will demonstrate the unique capabilities of the 3D-FAST technology. Rapid, imaging of improved voltage indicators will be paired with spatially-restricted light delivery for optogenetic neural modulation for the first time in a freely moving mouse. The optical imaging properties will be compared with ground-truth two-photon microscopy, and the functional consequence of neuromodulation will be dissected through behavioral assays. The 3D-FAST tool will bring novel capabilities to measuring and modulating large populations of neurons in freely-moving animals, to better understand the neural computations that underlie behavior. In addition, this body of work will lay the ground for future development of fully implantable optical recording and modulating units for use in freely-moving, untethered naturalistic behavior experiments.