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 该项目旨在开发体积钙成像二光子活动显微镜并将其商业化， vCAm™，一款革命性的新型 2 光子显微镜，基于称为光珠的技术突破 最近由 Alipasha Vaziri 博士及其同事（Lab. Neurotechnol. Biophys., Rockefeller Univ., New York, NY) vCAm 改变游戏规则的是执行能力。 在一个或多个细胞中几乎同时以细胞分辨率对单个神经元进行无与伦比的体内钙成像 小鼠大脑皮层的更多细胞结构区域，并且几乎同时在 30 个成像平面上 每个间隔约 16 µm（即总深度达 500 µm，包含 I-V 层），全帧速率至少为 12 赫兹。这些功能对于最终关联动物的刺激和/或行为状态至关重要。 离散地，以上下文相关的方式，与动物大脑中所有神经元的活动相关 参与这个过程，需要同时记录数十万个的活动 然而，当代的 2 光子显微镜却面临着多区域和多层神经元的问题。 神经科学研究人员需要记录之间的同时相互作用。 大脑的感觉、运动和视觉区域，但很难捕获如此大范围的活动 LBM 技术将成像速度的极限推向了极限。 通过消除连续激光脉冲之间的“死区时间”，当没有荧光时，荧光本身的物理性质 记录神经活动并同时进行扫描，是这种方法的唯一限制。 可以记录样本的速率是标签发出荧光所需的时间，这意味着大量的 与传统双光子显微镜捕获大脑的时间相同 其他技术，例如可以使用更少数量的脑细胞。 携带在自由活动的啮齿动物的头上，功能性磁共振成像，将电极插入 大脑或光纤光度测定法无法满足这一需求，该项目将改进原来的 LBM 发明。 基于在进行的试点工作，创建一个商业产品来传播这项重要的新技术。 Vaziri博士的实验室，显然vCAm将对神经科学领域产生重大影响 研究，包括针对中枢神经系统回路改变的推进研究 最终，这将与神经发育、神经精神和神经退行性疾病相关。 为开发针对各种复杂大脑的新治疗策略奠定了基础 在第一阶段，我们将通过开发原型硬件来证明这项新技术的可行性。 和软件；第二阶段的工作将侧重于创建 vCAm 的全部功能以供商业发布。 我们将密切对 vCAm 进行广泛的可行性研究、产品验证和可用性研究 与 Vaziri 博士的合作尚未实现商业化。