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.

项目摘要 大脑是建立在数十亿个神经连接的高度组织的3D层次结构上。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，神经通信的基本位。虽然最近出现的基因编码 电压指示器（GEVI）使得可以光学记录神经膜电压， 在利用细胞成像技术在千赫兹下成像毫米级体积电压成像方面存在着深刻的挑战 分辨率在这项提案中，我们的目标是通过开发一个单光子介观（即， 毫米级视场（FOV）体积电压成像，使用介观斜平面显微镜 （Meso-OPM）我们的技术将成像>1.8 mm 2 FOV，>0.1 mm深度穿透，1 KHz，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 Voltron与JF 525染料将用于我们提出的工作。Meso-OPM是光片显微镜的一种变体 (LSM)用单个主物镜透镜代替传统LSM中的两个。简化的光学设计 允许1）在LSM中利用高光子效率; 2）集成超快无源光学扫描以实现>1 MHz帧速率;以及3）灵活的光学设计，用于毫米FOV和蜂窝分辨率。除了有 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有效的深度学习框架来执行自学习。 有监督的4D去噪和神经元分割。该管道可实现10卷的海量数据处理 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 负责光流的运动补偿（即，当呈现漂移时保持身体位置 从眼睛一直到脊髓。总之，这项建议将大大提高我们的能力， 解剖大规模神经电路，以及人工神经电路的子系统建模和创建。