A robotic multi-armed two-photon microscope for imaging neural interactions across multiple brain areas

机器人多臂双光子显微镜，用于对多个大脑区域的神经相互作用进行成像

• 批准号: 10675439
• 负责人: MARK J SCHNITZER
• 金额: $ 79.54万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10675439
• 关键词: Achievement; Address; Adoption; Anatomy; Animal Behavior; Animals; Area; Articular Range of Motion; Articulation; Auditory area; BRAIN initiative; Basal Ganglia; Behavior; Brain; Brain region; Callithrix; Cells; Cerebellum; Cognitive; Color; Communities; Data; Development; Distal; Endowment; Feedback; Fluorescence; Freedom; Generations; Genetic; Head; Image; Imaging Device; Imaging Techniques; Individual; Institution; Intuition; Lateral Geniculate Body; Lighting; Location; Mechanics; Methods; Microscope; Microscopy; Monitor; Motion; Motor; Motor Cortex; Mus; Neurons; Neurosciences; Octopus; Operative Surgical Procedures; Optics; Performance; Population; Positioning Attribute; Preparation; Primates; Pulvinar structure; Readiness; Research; Robot; Robotics; Rodent; Role; Rotation; Route; Scanning; Sensory; Shipping; Site; System; Techniques; Technology; Thalamic structure; Tissues; Visible Radiation; Visual; Visual Cortex; Work; arm; arm movement; awake; cell type; design; design,build,test; dexterity; flexibility; imaging modality; imaging study; imaging system; improved; invention; kinematics; lens; manufacture; meter; microendoscope; microscopic imaging; millimeter; miniaturize; motor behavior; neural; neuroimaging; new technology; open source; optical imaging; optogenetics; restraint; superior colliculus Corpora quadrigemina; tool; two-photon; usability

Abstract Among the BRAIN Initiative’s most important achievements are the genetic identification of many new neurons- types and the creation of genetic tools to access these cell types. However, uncovering the functional roles of these neuron types and how they cooperate across brain areas to generate mammalian behavior remains an outstanding challenge. Thus, inventing ways to monitor how large populations of genetically identified neurons interact across multiple regions of the brain is crucial if we are to comprehend global brain dynamics. Today, electrical recording methods can track neural activity across multiple areas but cannot easily target neurons of specific types. Widefield and two-photon mesoscopes can image the dynamics of identified neuron-types across millimeter-scale regions of cortical tissue but cannot access the distributed sets of cortical and subcortical regions that comprise the major nodes of the brain’s sensory, cognitive, or motor circuits. To clear this impasse, we invented the ‘Octopus’, a robotic imaging system with multiple articulated optical arms, each a two-photon microscope, that can be flexibly positioned around the brain to record neural activity concurrently in multiple superficial or deep areas of a head-restrained behaving rodent or primate. We designed, built, and tested an initial version of the Octopus with 4 arms, each of which has 5 mechanical degrees of freedom and a micro-optic probe at its tip for two-photon imaging. The design of the arms is based on ideas from surgical robotics and uses remote center-of-motion kinematics to provide a versatile repertoire of robotic arm movements. Using this system, a visual neuroscientist can concurrently image neural activity in the lateral geniculate nucleus, visual cortex, superior colliculus, and pulvinar, and a motor neurophysiologist can image activity in the motor cortex, basal ganglia, cerebellum, and motor thalamus. In this project, we will enhance the optical and mechanical design of each Octopus arm and prepare the system for wide dissemination through open-source and commercial routes. Each arm will gain the optical functionality of a state-of-the-art, two-photon microscope for imaging large-scale neural ensemble activity. Specifically, each arm will incorporate optogenetics and allow dual-color two-photon imaging over an 800-µm- wide field of view. These capabilities will allow neuroscientists to monitor two genetically identified neuron- types in each of 4 brain areas, to perturb the dynamics of these cells with optogenetics, and to observe the effects of these manipulations on animal behavior and activity in the other 3 areas. We will also streamline the mechanical design to simplify the initial assembly of the Octopus for new users and to endow the robot arms with additional dexterity. The new design will also be motorized and will provide users with highly intuitive means of precisely steering the robot arms. Finally, to iteratively improve the performance and usability of the Octopus and to validate its readiness for dissemination as a groundbreaking new technology, we will work closely with 7 beta-tester labs to implement multi-area neural imaging studies in awake behaving mice and marmosets.

抽象的 BRAIN Initiative 最重要的成就之一是对许多新神经元类型进行了遗传鉴定，并创建了访问这些细胞类型的遗传工具，揭示了这些神经元类型的功能作用以及它们如何跨大脑区域合作以产生哺乳动物行为。因此，如果我们要理解全球大脑动态，发明方法来监测大量经过基因识别的神经元如何在大脑的多个区域相互作用至关重要。如今，电记录方法可以跟踪多个区域的神经活动，但无法追踪。轻松针对特定类型的神经元。宽场和双光子介观镜可以对皮质组织毫米级区域内已识别神经元类型的动态进行成像，但无法访问构成大脑感觉、认知或运动回路主要节点的皮质和皮质下区域的分布式集合。为了打破这一僵局，我们发明了“章鱼”，这是一种机器人成像系统，具有多个铰接光学臂，每个光学臂都有一个双光子显微镜，可以灵活地定位在大脑周围，同时记录神经活动。我们设计、建造并测试了具有 4 个手臂的章鱼的初始版本，每个手臂都有 5 个机械自由度，并在其尖端有一个微光学探针。用于双光子成像的手臂的设计基于外科机器人学的想法，并使用远程运动中心运动学来提供机器人手臂运动的多种功能，视觉神经科学家可以同时使用该系统。对外侧膝状核、视觉皮层、上丘和枕丘的神经活动进行成像，运动神经生理学家可以对运动皮层、基底神经节、小脑和运动丘脑的活动进行成像。在这个项目中，我们将增强光学和机械功能。每个章鱼臂的设计，并为通过开源和商业途径广泛传播该系统做好准备，每个臂都将获得最先进的双光子显微镜的光学功能，用于成像。具体来说，每个手臂都将结合光遗传学，并允许在 800 微米宽的视野内进行双色双光子成像，这些功能将使神经科学家能够监测每个手臂中两种基因识别的神经元类型。 4 个大脑区域，通过光遗传学扰乱这些细胞的动态，并观察这些操作对其他 3 个区域的动物行为和活动的影响。我们还将简化机械设计，以简化初始组装。为新用户设计的章鱼，赋予机器人手臂额外的灵活性。新设计还将实现机动化，为用户提供高度直观的精确操控机器人手臂的方法，最终不断提高章鱼的性能和可用性。为了验证其作为一项突破性新技术的传播准备，我们将与 7 个 beta 测试实验室密切合作，在清醒行为的小鼠和狨猴中实施多区域神经成像研究。