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

摘要 该项目旨在开发2 P-ActivityScope ™，这是一种革命性的新型显微镜， 第二代扫描线角投影（SLAP 2）双光子激光扫描 显微镜，最近开发的卡斯帕Podgorski博士（珍妮研究校园，霍华德休斯 Medical Institute，Ashburn，VA）。2 P-ActivityScope中改变游戏规则的创新是能够执行 （i）以亚细胞空间分辨率对神经元群体中的神经元活动进行光学成像， 使用多种荧光指示剂（例如， 电压，[神经递质]）以及（ii）皮质柱的高分辨率体积钙成像 在40 Hz的频率下，这些能力对于最终解码信息是如何表示的至关重要 并由数十亿个密集互连的神经元处理， 系统现代的双光子显微镜，功能性磁共振成像， 将电极插入大脑，或者纤维光度法可以满足这种需要。该项目将传播和改进 最初的实验室制造的SLAP 2发明，为这项重要的新技术创造了一个商业产品。基于 在Janelia进行的试点工作中，很明显，2 P-ActivityScope将对该领域产生重大影响 神经科学研究，包括推进研究集中在中枢神经系统电路的变化与 神经发育、神经精神和神经变性病症。最终，这将导致 为开发针对各种复杂脑部疾病的新型治疗策略提供了更好的基础。在 第一阶段，我们将通过开发原型硬件和软件来证明这项新技术的可行性; 第二阶段的工作将集中于为商业发布创建2 P-ActivityScope的全部功能。我们 将进行广泛的可行性研究，产品验证和可用性研究的2 P-ActivityScope在关闭 与Podgorski博士和我们的学术合作伙伴合作。一项竞争性技术 市售。

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.