BRAIN EAGER: Three Dimensional Optical Control of Neuronal Circuits during Behavior

系统神经科学的一个中心目标是根据控制行为的神经元回路来描述行为。这在哺乳动物的大脑中构成了一个巨大的挑战，因为行为被认为依赖于广泛分布的神经表征，这在技术上很难在大尺度上监测或在细胞分辨率上操纵。这项研究将通过在线存储库和研讨会实现、优化并最终广泛传播一种三维双光子显微镜，利用模式照明策略来快速操纵和监测行为动物的大型神经元群。首先，PI将实现并优化双光刺激系统，使用数字微镜设备实现宽视场的精确二维照明，以及使用数字全息技术实现高分辨率三维刺激。光刺激将与双光子共振扫描成像相结合，以实现大脑容量中数百个神经元的快速采样率。其次，这些技术将被证明在体内使用，在头部固定行为的啮齿动物中，优化其实验应用的适用性。这种方法将使动态测试任意神经元的功能角色和行为啮齿类动物的选择组合成为可能。该项目将开发和实施创新的光学方法，用于神经系统和其他可兴奋的生物组织的精细图案刺激。该方法将利用现有的光敏离子通道，利用光遗传学技术将其插入神经元中。这种有计划的光学照明方法将首次允许对大脑活动进行精细的空间控制和监测。此外，这些发展将为任何光学可及组织的细胞内和细胞间信号过程的高分辨率三维光学控制提供新的应用，大大拓宽了它们在生物研究和生物工程中的适用性。该项目将使所有开发出来的方法在感兴趣的科学家之间广泛传播。此外，研究结果将用于提高公众对应用光学与日常生活的相关性的认识。这些目标将通过协调一致的外展计划来实现，包括在冷泉港实验室开设光学成像课程、志愿实习以及在长岛和纽约市的当地学校授课。

A central goal of systems neuroscience is to describe behaviors in terms of the neuronal circuits that control them. This constitutes a monumental challenge in the mammalian brain because behaviors are thought to rely on widely distributed neural representations, which are technically difficult to monitor at large scales or to manipulate at cellular resolution. This research will implement, optimize, and ultimately broadly distribute via online repositories and workshops, a three-dimensional, two-photon microscope, exploiting patterned illumination strategies for fast manipulation and monitoring of large neuronal populations in behaving animals. First, the PI will implement and optimize a dual photo-stimulation system to enable both precise two-dimensional illumination across wide fields of view using a digital micro-mirror device, as well as high-resolution three-dimensional stimulation using digital holography. Photo-stimulation will be combined with two photon resonant scanning imaging to achieve fast sampling rate of hundreds of neurons across large brain volumes. Second, these technologies will be demonstrated for in vivo use, in head-fixed behaving rodents, optimizing their applicability for experimental use. This approach will enable dynamic testing of the functional roles of arbitrary neurons and combinations of choice in behaving rodents. This project will develop and implement innovative optical methods for fine patterned stimulation of nervous systems and other excitable biological tissues. The approach will exploit current light-sensitive ion channels that can be inserted into neurons using the techniques of optogenetics. The planed optical illumination methods will, for the first time, allow fine spatial control and monitoring of brain activity. Further, these developments will provide new applications for high-resolution three-dimensional optical control of intra- and inter-cellullar signaling processes in any optically accessible tissue, greatly broaden their applicability in biological research and bioengineering. The project will result in broad distribution among interested scientists of all the methods developed. In addition, the results will be used to increase the awareness among the general public of the relevance of applied optics to daily life. These goals will be achieved through a concerted outreach program including optical imaging courses in Cold Spring Harbor Laboratory, volunteering internships, and lectures to local schools in Long Island and New York City.