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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

基本信息

批准号：
9770567
负责人：
Emily Gibson
金额：
$ 80.09万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-30 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9770567
关键词：
3-Dimensional 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 Intelligence Label Lasers Lateral Light Lighting Location Microscope 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 TimeLine Tissues Validation Weight Work awake base brain tissue calcium indicator confocal imaging 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 microscopy two-photon vocal learning

项目摘要

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.

这是响应 RFA-NS-17-00 4“BRAIN Initiative：变革性优化”的应用程序神经系统大规模记录和调制技术 (U01)”。概念验证在该出版物中，我们展示了一种新型光纤耦合微型显微镜，可实现 3D 共焦成像使用电润湿透镜技术进行轴向聚焦，荧光标记脑组织中的神经元。这个低体重仪（<2 g）适合手术植入，实现全光学唤醒行为记录在小鼠大脑深处进行轴向聚焦，与商用设备相比这是一个重大进步。在这项提案中，我们提出了一项重大突破，展示了通过光纤耦合的成功成像显微镜使用双光子显微镜。利用这一结果，我们建议优化我们的证明概念 E-FCM（电润湿光纤耦合显微镜），使其兼容双光子激发 (2PE-FCM) 适用于记录和调节清醒行为小鼠的神经活动。我们的 2PE-FCM 将将全息双光子视蛋白刺激与空间光调制器 (SLM) 相结合，以实现快速 3D 体积双光子成像和神经元活动的调节。 2PE-FCM 的优化将是通过测试版用户与物理学家、工程师和多学科小组的定期互动来实现神经科学家在设计实施和测试设备的并行过程中。 2PE-FCM 测试将采用分步方法，旨在有效生成适合研究本地的设备不同物种的不同大脑区域的回路。我们与以下机构合作提出了可靠的传播计划智能成像创新公司将该设备商业化。 2PE-FCM 的验证将是一项重大工作神经回路全光学询问的进展为理解神经基础创造了新的场所的行为。目标 1. 技术方法：光遗传学激活和同时读出深部使用带有可变焦电润湿透镜的新型光纤耦合显微镜的脑神经元不使用移动部件。目标 2. 里程碑和时间表：验证并传播 2PE-FCM 设备研究自由行为动物行为的神经基础。