Controlled neuronal firing in vivo using two photon spatially shaped optogenetics

使用两个光子空间形状光遗传学控制体内神经元放电

• 批准号: 9404641
• 负责人: Emily Gibson
• 金额: $ 70.88万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9404641
• 关键词: Animal Model; Animals; Area; BRAIN initiative; Behavior; Brain; Brain imaging; Brain region; Calcium; Caliber; Cell Culture Techniques; Cells; Coupled; Custom; Data; Decision Making; Devices; Disease; Engineering; Fiber; Generations; Head; Health; Hippocampus (Brain); Image; Individual; Label; Lasers; Lateral; Light; Lighting; Location; Microscope; Microscopy; Movement; Mus; Nervous system structure; Neurons; Neurophysiology - biologic function; Operative Surgical Procedures; Opsin; Optics; Pattern; Phenotype; Phototoxicity; Physiologic pulse; Process; Publications; Resolution; Site; Solid; Songbirds; System; Technology; Testing; Three-Dimensional Imaging; Tissues; Validation; Weight; Work; awake; base; brain tissue; calcium indicator; design; flexibility; fluorescence imaging; fluorescence microscope; implantation; improved; in vivo; innovation; instrument; interest; lens; light scattering; light weight; meetings; mind control; miniaturize; multi-electrode arrays; multidisciplinary; myelination; neural circuit; nonhuman primate; novel; olfactory bulb; optical fiber; optical imaging; optogenetics; photoactivation; relating to nervous system; response; sensor; sensory input; social communication; two-photon; vocal learning

This is an application in response to RFA-NS-17-00 4 “BRAIN Initiative: Optimization of Transformative Technologies for Large Scale Recording and Modulation in the Nervous System (U01)”. In a proof-of-concept publication we demonstrated a novel fiber-coupled miniature microscope that achieved 3D confocal imaging of fluorescently labeled neurons in brain tissue using electrowetting lens technology for axial focusing. This low weight instrument (<2 g) is suitable for surgical implantation to achieve all optical awake behaving recording with axial focusing deep in the mouse brain, a significant advance compared to commercially available devices. In this proposal we present a major breakthrough demonstrating successful imaging through the fiber coupled microscope using two-photon microscopy. Leveraging on this result we propose to optimize our proof of concept E-FCM (electrowetting-fiber coupled microscope) by making it compatible for two-photon excitation (2PE-FCM) suitable for recording and modulating neural activity in awake behaving mice. Our 2PE-FCM will incorporate holographic two-photon opsin stimulation with a spatial light modulator (SLM) for rapid 3D volumetric two-photon imaging and modulation of neuronal activity. Optimization of the 2PE-FCM will be achieved by regular interactions of beta users with a multidisciplinary group of physicists, engineers and neuroscientists in a parallel process of design implementation and testing the device. Testing of the 2PE-FCM will employ a step-by-step approach designed for effective generation of a device suitable for studying local circuits in different brain areas in different species. We propose a solid dissemination plan partnering with Intelligent Imaging Innovations Inc. to commercialize the device. Validation of the 2PE-FCM will be a major advance in all-optical interrogation of neural circuits creating a new venue for understanding the neural basis of behavior. Aim 1. TECHNICAL APPROACH: Optogenetic activation and simultaneous readout of deep brain neurons using a novel fiber coupled microscope with a variable focus electrowetting lens using no moving parts. Aim 2. MILESTONES AND TIMELINE: Validate and disseminate the 2PE-FCM device for studies of the neural basis of behavior in freely behaving animals.

这是一个响应RFA-NS-17-00 4“BRAIN Initiative: Optimization of Transformative”的应用