System for Volumetric 2-photon Imaging of Neuroactivity Using Light Beads Microscopy

使用光珠显微镜对神经活动进行体积 2 光子成像的系统

• 批准号: 10755027
• 负责人: JACOB R GLASER
• 金额: $ 99.98万
• 依托单位: MICROBRIGHTFIELD, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-05 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10755027
• 关键词: System; Volumetric; photon; Imaging; Neuroactivity

Abstract This project aims to develop and commercialize the Volumetric Calcium Imaging 2-Photon Activity Microscope, vCAm™, a revolutionary new 2-photon microscope based on a technological breakthrough called Light Beads Microscopy (LBM) that was recently developed by Dr. Alipasha Vaziri and co-workers (Lab. Neurotechnol. Biophys., Rockefeller Univ., New York, NY). The game-changing innovation in the vCAm is the ability to perform unparalleled in vivo calcium imaging of individual neurons at cellular resolution nearly simultaneously in one or more cytoarchitectonic regions of the mouse cerebral cortex, and nearly simultaneously in 30 imaging planes each ~16 µm apart (i.e., up to a total depth of 500 µm, encompassing layers I-V) at a full-frame rate of at least 12 Hertz. These capabilities are crucial for ultimately correlating stimuli and/or behavioral states of an animal discretely, in a context-dependent manner, with the activity of all neurons in the brain of the animal that are involved in this process, which requires simultaneous recording of the activity of hundreds of thousands of neurons in a multi-regional and multi-layer manner. However, contemporary 2-photon microscopy suffers from a fundamental limitation. Neuroscience researchers need to record simultaneous interactions between the sensory, motor and visual regions of the brain, but it is difficult to capture the activity in such a broad volume of the brain without sacrificing resolution or speed. The LBM technology pushes the limits of imaging speed to the physical nature of fluorescence itself by eliminating the “dead time” between sequential laser pulses when no neuroactivity is recorded and at the same time the need for scanning. With this approach, the only limit to the rate at which samples can be recorded is the time that it takes the tags to fluoresce, meaning wide volumes of the brain can be recorded within the same time it would take a conventional two-photon microscope to capture a much smaller number of brain cells. Other technology, such as miniaturized 2-photon microscopes that can be carried on the head of freely moving rodents, functional magnetic resonance imaging, inserting electrodes into the brain, or fiber photometry do not fulfill this need. This project will improve upon the original LBM invention to create a commercial product for disseminating this important new technology. Based on pilot work performed at Dr. Vaziri's laboratory, it is clear that the vCAm will make a significant impact on the field of neuroscience research, including advancing studies focused on alterations in the circuitry of the central nervous system 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 the feasibility of this novel technology by developing prototype hardware and software; work in Phase II will focus on creating the full functionality of the vCAm for commercial release. We will perform extensive feasibility studies, product validation and usability studies of the vCAm in close collaboration with Dr. Vaziri. A competing technology is not commercially available.

抽象的 该项目旨在开发和商业化体积钙成像2光子活性显微镜， VCAM™，一种革命性的新2光子显微镜，基于一个名为Light Beads的技术突破 显微镜（LBM）最近由Alipasha Vaziri博士和同事（Lab。Neurotechnol）开发。 Biophys，洛克菲勒大学，纽约，纽约）。 VCAM中改变游戏规则的创新是表演的能力 几乎同时在一个或 小鼠大脑皮层的更多细胞结构区域，几乎同时在30个成像平面 每个〜16 µm相距（即总深度为500 µm，包含I-V层），至少为全帧速率 12赫兹。这些能力对于最终将动物的刺激和/或行为状态关联至关重要 以上下文依赖性的方式离散地具有动物大脑中所有神经元的活性 参与此过程，这需要简单记录数十万的活动 以多区域和多层方式进行神经元。但是，现代2光子显微镜遭受 基本限制。神经科学研究人员需要记录 大脑的感觉，运动和视觉区域，但是很难捕获如此广泛的活动 大脑而无需牺牲解决方案或速度。 LBM技术将成像速度的极限推向 荧光本身的物理性质，通过消除否时，消除顺序激光脉冲之间的“死时间” 记录神经活性，同时进行扫描。使用这种方法，唯一的限制是 可以记录样品的速率是将标签带到荧光的时间，这意味着很大的体积 可以在传统的两光子显微镜捕获的同时记录大脑 脑细胞数量少得多。其他技术，例如微型2光子显微镜，可以是 自由移动啮齿动物的头部，功能磁共振成像，将电子插入 大脑或光纤光度法无法满足这种需求。该项目将在原始LBM事故中改善 创建用于传播这项重要新技术的商业产品。基于执行的飞行员工作 Vaziri博士的实验室，很明显，VCAM将对神经科学领域产生重大影响 研究，包括前进的研究，重点是中枢神经系统电路的变化 与神经发育，神经精神病学和神经退行性疾病有关。最终，这将是 为开发各种复杂大脑的新型治疗策略而改善了基础 疾病。在第一阶段，我们将通过开发原型硬件来证明这种新技术的可行性 和软件；第二阶段的工作将着重于创建VCAM的全部功能，以供商业发布。 我们将对VCAM进行广泛的可行性研究，产品验证和可用性研究 与Vaziri博士的合作。竞争技术无法商业上使用。