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

项目摘要 为了进一步了解神经回路的功能，我们需要新的工具， 从涉及共同神经元的大量神经元中收集同时测量值。 计算并提供精确的功能调制。清醒动物的光学成像 钙或电压指示器提供来自个体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的视场， 包括单个大脑区域的视图，以及头部固定的要求，这防止了 自然主义行为。此外，大多数光学成像系统不允许同时高- 分辨率功能成像与空间定位光遗传学调节的组合。 为了应对这一挑战，我们提出开发一种光学设备（“3D-FAST”），其允许快速、真实的- 时间体积神经记录和精确的光学刺激。通过配对微型阵列 微图案化LED发射器与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，可定制的透镜阵列， 允许记录大规模双向神经接口，用于神经网络的闭环调制， 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 Hz成像，以及3D-FAST-GEVI，其将提供1 kHz的帧率， 单平面结合较慢的高分辨率体积扫描。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 ASAP和Voltron，最后结合局部光遗传学进行闭环反馈。 总之，这些实验将展示3D-FAST技术的独特功能。迅速， 改进的电压指示器的成像将与空间受限的光传输配对， 光遗传学神经调制的第一次在一个自由移动的小鼠。光学成像特性 将与地面实况双光子显微镜进行比较，以及 神经调节将通过行为分析来剖析。3D-FAST工具将带来新颖的 在自由移动的动物中测量和调节大量神经元的能力， 理解行为背后的神经计算。此外，这项工作将奠定 为未来开发完全植入式光学记录和调制装置奠定了基础， 自由移动，不受束缚的自然行为实验

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.