Mesoscopic microscopy for ultra-high speed and large-scale volumetric brain imaging

用于超高速和大规模脑体积成像的介观显微镜

• 批准号: 10634911
• 负责人: Ji Yi
• 金额: $ 53.87万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10634911
• 关键词: 3-Dimensional; Action Potentials; Address; Adopted; Algorithms; Animals; Area; Behavior; Biological; Bite; Brain; Brain imaging; Budgets; Central Nervous System; Code; Collection; Communication; Compensation; Data; Data Set; Detection; Development; Dyes; Electronics; Eye; Fishes; Fluorescence; Generations; Goals; Image; Kinetics; Label; Larva; Lasers; Light; Maps; Mechanics; Membrane; Membrane Potentials; Methods; Microscopy; Modeling; Motion; Motor; Mus; Nervous System; Neurons; Neurosciences; Noise; Optics; Pattern; Penetration; Performance; Photobleaching; Photons; Photoreceptors; Posture; Resolution; Retina; Sampling; Scanning; Signal Transduction; Speed; Spinal Cord; Stimulus; Structure; System; Techniques; Tectum Mesencephali; Testing; Three-Dimensional Imaging; Time; Tissues; Training; Variant; Work; Zebrafish; body position; brain volume; computerized data processing; data analysis pipeline; deep learning; deep learning algorithm; deep learning model; denoising; design; detector; flexibility; imaging system; improved; instrumentation; lens; millimeter; motor behavior; nervous system imaging; network architecture; neural; neural circuit; neuroimaging; optic flow; processing speed; prototype; response; spatiotemporal; supervised learning; three-dimensional modeling; transmission process; visual stimulus; voltage

PROJECT SUMMARY The brain is built on billions of neural connections in a highly organized 3D hierarchy. At the same time, neural activity is highly dynamics that requires kilohertz imaging rate to capture action potentials and sub-threshold voltage signals, the fundamental bit for neural communication. While the recent advent of genetically encoded voltage indicators (GEVIs) makes it possible to optically record the neural membrane voltage, the technical challenges are profound in imaging millimeter-scale volumetric voltage imaging at kilohertz with cellular resolution. In this proposal, we aim to address the challenges by developing a one-photon mesoscopic (i.e. millimeter scale field of view, FOV) volumetric voltage imaging, using mesoscopic oblique plane microscopy (Meso-OPM). Our technique will image >1.8 mm2 FOV, >0.1 mm depth penetration at 1 KHz, capable of recording voltage signals across an entire nervous system of a Zebrafish larva. The bright and stable GEVIs Voltron with JF525 dye will be used in our proposed work. Meso-OPM is a variant of light sheet microscopy (LSM), with a single primary objective lens instead of two in conventional LSM. The simplified optical design allows 1) leveraging high photon efficiency in LSM; 2) integrating ultra-fast passive optical scanning to achieve >1 MHz frame rate; and 3) flexible optical designs for millimeter FOV and cellular resolution. In addition to the technical challenges for large-scale ultrafast 3D imaging, the effective data processing pipeline for massive data is also highly desirable. To this end, we propose a robust and efficient deep learning framework to perform self- supervised 4D denoising and neuron segmentation. The pipeline enable massive data processing at 10 volume per second for the downstream neuroscience studies. Finally, to demonstrate the utility of proposed techniques, we will image Zebrafish in response to optic flow by a drifting grating visual stimuli. We will identify neural circuitry responsible to the motion compensation to the optic flow (i.e. maintaining body position when presented drifting grating) from eyes all the way to spinal cord. Altogether, this proposal will greatly improve our capability of dissecting large-scale neural circuitry, and the sub-sequent modeling and creation of artificial neural circuits.

项目概要 大脑是建立在高度组织的 3D 层次结构中的数十亿个神经连接之上的。与此同时，神经 活动是高度动态的，需要千赫兹成像速率来捕获动作电位和亚阈值 电压信号，神经通讯的基本位。虽然最近出现了基因编码 电压指示器（GEVI）使得光学记录神经膜电压成为可能，该技术是 利用蜂窝技术对千赫兹毫米级体积电压成像进行成像面临着严峻的挑战 解决。在本提案中，我们的目标是通过开发单光子介观（即， 毫米级视场（FOV）体积电压成像，使用介观斜平面显微镜 （细观-OPM）。我们的技术将在 1 KHz 下成像 >1.8 mm2 FOV、>0.1 mm 深度穿透，能够 记录斑马鱼幼虫整个神经系统的电压信号。明亮稳定的GEVI 我们提议的工作中将使用带有 JF525 染料的 Voltron。 Meso-OPM 是光片显微镜的一种变体 (LSM)，具有单个主物镜，而不是传统 LSM 中的两个。简化的光学设计 允许 1) 利用 LSM 中的高光子效率； 2）集成超快无源光学扫描，实现>1 MHz 帧速率； 3) 针对毫米级视场和蜂窝分辨率的灵活光学设计。除了 大规模超快3D成像的技术挑战，海量数据的有效数据处理管道 也是非常理想的。为此，我们提出了一个强大而高效的深度学习框架来执行自我学习 监督 4D 去噪和神经元分割。该管道可实现 10 卷的海量数据处理 每秒用于下游神经科学研究。最后，为了证明所提出的技术的实用性， 我们将通过漂移光栅视觉刺激来成像斑马鱼对光流的响应。我们将识别神经回路 负责对光流的运动补偿（即在出现漂移时保持身体位置） 光栅）从眼睛一直到脊髓。总而言之，这个建议将大大提高我们的能力 剖析大规模神经回路，以及人工神经回路的后续建模和创建。