Microscope system for large scale optical imaging of neuronal activity using kilohertz frame rates

摘要 该项目旨在开发2P-ActivityScope™，这是一种革命性的新显微镜，基于一种技术 称为第二代扫描线角度投影(SLAP2)双光子激光扫描的突破 显微镜是由Kaspar Podgorski博士(Janelia Research Campus，Howard Hughes)最近开发的 弗吉尼亚州阿什伯恩医学院)。2P-ActivityScope中改变游戏规则的创新是执行 无与伦比的(I)亚细胞空间分辨率的神经元群体中神经元活动的光学成像和 在体内和体外使用各种荧光指示剂(例如 电压，[神经递质])以及(Ii)皮质柱的高分辨率体积钙成像。 活体内40赫兹的神经元。这些功能对于最终解码信息的表示方式至关重要 并由组成哺乳动物中枢神经的数十亿个紧密相连的神经元进行处理 系统。既不是当代的双光子显微镜，功能磁共振成像，插入 电极进入大脑，或纤维光度测量可以满足这一需求。该项目将传播和改进 原创的实验室建造的SLAP2发明，为这项重要的新技术创造了商业产品。基座 关于在Janelia开展的试点工作，2P-ActivityScope显然将对实地产生重大影响 神经科学研究的进展，包括重点研究中枢神经系统相关回路的改变 神经发育、神经精神和神经退行性疾病。最终，这将导致 改进了为广泛的复杂脑部疾病开发新的治疗策略的基础。在……里面 第一阶段，我们将通过开发原型硬件和软件来论证这一新技术的可行性； 第二阶段的工作将集中于创建2P-ActivityScope的全部功能，用于商业发布。我们 将对2P-ActivityScope进行广泛的可行性研究、产品验证和可用性研究 与波德戈尔斯基博士和我们的学术合作伙伴的合作。一项与之竞争的技术不是 商业上可以买到的。

Abstract This project aims to develop the 2P-ActivityScope™, a revolutionary new microscope based on a technological breakthrough called second-generation Scanned Line Angular Projection (SLAP2) two photon laser scanning microscopy that was recently developed by Dr. Kaspar Podgorski (Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA). The game-changing innovation in the 2P-ActivityScope is the ability to perform unparalleled (i) optical imaging of neuronal activity in populations of neurons at subcellular spatial resolution and temporal resolution in the millisecond range both in vivo and in vitro using a variety of fluorescent indicators (e.g. voltage, [neurotransmitter]) as well as (ii) high-resolution volumetric calcium imaging of columns of cortical neurons at 40 Hz in vivo. These capabilities are crucial for ultimately decoding how information is represented and processed by the billions of densely interconnected neurons comprising the mammalian central nervous system. Neither contemporary 2-photon microscopes, functional magnetic resonance imaging, inserting electrodes into the brain, or fiber photometry can serve this need. This project will disseminate and improve upon the original, lab-built SLAP2 invention to create a commercial product for this important new technology. Based on pilot work performed at Janelia, it is clear that the 2P-ActivityScope will make a significant impact on the field of neuroscience research, including advancing studies focused on alterations in CNS circuitry associated with neurodevelopmental, neuropsychiatric and neurodegenerative disorders. Ultimately, this will result in an improved basis for developing novel treatment strategies for a wide spectrum of complex brain diseases. In Phase I we will demonstrate feasibility of this novel technology by developing prototype hardware and software; work in Phase II will focus on creating the full functionality of the 2P-ActivityScope for commercial release. We will perform extensive feasibility studies, product validation and usability studies of the 2P-ActivityScope in close collaboration with Dr. Podgorski and our academic collaboration partners. A competing technology is not commercially available.