High-speed volumetric imaging of neural activity throughout the living brain

整个活体大脑神经活动的高速体积成像

• 批准号: 9404832
• 负责人: NA Ji
• 金额: $ 89.32万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-15 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9404832
• 关键词: Accelerometer; Adopted; Adoption; Algorithms; Animal Model; Animals; Axon; Biology; Brain; Brain imaging; Calcium; Cell Nucleus; Complex; Computer software; Corpus striatum structure; Data; Data Analyses; Data Science; Dendrites; Dendritic Spines; Dimensions; Ensure; Event; Ferrets; Fluorescence; Fluorescence Microscopy; Goals; Head; Hypothalamic structure; Image; Imaging technology; Individual; Label; Laboratories; Lateral; Measures; Methods; Microscope; Monitor; Morphologic artifacts; Motion; Motor; Mus; Neurobiology; Neurons; Neurosciences Research; Optics; Output; Photons; Population; Resistance; Resolution; Scanning; Signal Transduction; Speed; Structure; Synapses; System; Techniques; Technology; Thick; Three-Dimensional Imaging; Time; Tissue imaging; Work; Zebrafish; adaptive optics; base; brain tissue; brain volume; experience; fluorescence microscope; fly; functional plasticity; imaging modality; in vivo; light scattering; microendoscopy; nervous system disorder; neural circuit; novel strategies; operation; prevent; relating to nervous system; sensory input; spatiotemporal; temporal measurement; tool; two-photon

To understand how the brain computes, we need to understand how individual neurons in a circuit integrate their numerous inputs into output signals, as well as how they work together to encode a sensory input or execute a motor command in a behaving animal. Circuits and neurons are three-dimensional (3D) and can extend over hundreds or thousands of microns. Therefore, understanding their operations requires monitoring their activity at both synaptic and cellular resolution in 3D at image rates that capture all activity events. Behaving animals present a host of challenges to this goal. Existing 3D imaging technologies suffer from insufficient volume imaging speed, brain-motion-induced image artifacts, as well as complex hardware and software implementation. These limitations have prevented their adoption by biology laboratories and remain a technical barrier for neuroscience research. Successful completion of our proposal will overcome these limitations and profoundly impact neuroscience research. We recently developed a Bessel focus scanning technology (BEST) that is easily integrated into existing two-photon microscopes, resistant to motion artifacts, and have already achieved 30-Hz, synapse-resolving volumetric imaging of sparsely labelled neuronal populations in a wide variety of model organisms. In this proposal, combining the expertise of microscopists, biologists, and data scientists, we propose to further optimize BEST to enable high-speed, high-throughput, and high-resolution volumetric activity recording of both sparsely and densely labelled circuits throughout the living brain. We aim to record whole-brain activity in the fly at >10 Hz and through-cortex volume imaging in the mouse at ~2Hz. By combining BEST with microendoscopy, we aim to achieve synaptic-resolution volumetric microendoscopic imaging at 30 Hz and use it to study structural and functional plasticity in deeply buried nuclei of the mouse brain. By correcting brain-induced optical aberrations, adaptive optics will enable BEST to maintain synapse resolution throughout the entire mouse cortex. Easily adoptable, BEST has already been integrated into multiple two-photon fluorescence microscopes in laboratories worldwide. With a continuously expanding user base, the proposed optimization project will immediately benefit a wide range of laboratories, allowing them to study volumetric neural activity at unprecedented high spatiotemporal resolution throughout the living brain.

要了解大脑如何计算，我们需要了解电路中的各个神经元如何集成 它们的大量输入转化为输出信号，以及它们如何协同工作来编码感官输入或 在行为动物中执行运动命令。电路和神经元是三维 (3D) 的，可以 延伸超过数百或数千微米。因此，了解他们的运作需要 以捕获所有活动的图像速率以 3D 方式监控突触和细胞分辨率的活动 事件。行为动物对这一目标提出了许多挑战。现有 3D 成像技术受到影响 由于体积成像速度不足、大脑运动引起的图像伪影以及复杂的 硬件和软件实现。这些限制阻碍了它们被生物学采用 实验室，仍然是神经科学研究的技术障碍。成功完成我们的提案 将克服这些限制并深刻影响神经科学研究。我们最近开发了一个 贝塞尔聚焦扫描技术（BEST）可轻松集成到现有的双光子显微镜中， 抗运动伪影，并且已经实现了 30 Hz、突触解析的体积成像 各种模型生物中稀疏标记的神经元群体。在该提案中，结合 结合显微镜学家、生物学家和数据科学家的专业知识，我们建议进一步优化 BEST 实现稀疏和高分辨率的体积活动记录 整个活体大脑中密集标记的电路。我们的目标是在 >10 时记录果蝇的全脑活动 Hz 和小鼠中约 2Hz 的穿过皮层体积成像。通过将 BEST 与显微内窥镜相结合， 我们的目标是在 30 Hz 下实现突触分辨率体积显微内窥镜成像，并用它来研究 小鼠大脑深埋核的结构和功能可塑性。通过纠正大脑诱发的 光学像差，自适应光学将使 BEST 在整个过程中保持突触分辨率 小鼠皮质。易于采用，BEST 已集成到多个双光子荧光中 世界各地实验室的显微镜。随着用户群的不断扩大，建议 优化项目将立即使广泛的实验室受益，使他们能够进行研究 在整个活体大脑中以前所未有的高时空分辨率进行体积神经活动。