Next generation in vivo miniature microscopes integrating dual-color imaging, 3D imaging, and optogenetic stimulation with a cloud-compatible data acquisition platform

项目摘要 我们对神经精神障碍的理解正在从简单的模型转向 强调神经递质失衡，转向基于神经异常的更复杂的理论 电路功能。这对于了解精神分裂症和创伤后应激障碍等障碍尤为重要 无视简单的生物化学解释，但现在已知涉及神经集合的异常模式 活动。即使对于其他类型的大脑疾病，例如神经退行性疾病(例如帕金森氏病)， 或有分子遗传、细胞或环境原因的神经发育障碍(如自闭症)， 越来越多的人意识到，这种功能障碍最终表现在神经回路水平上。 大脑，并表现为神经回路动力学异常。然而，我们仍然缺乏至关重要的基本理解 正常行为期间的神经活动模式，以及这些模式在疾病中是如何改变的。许多啮齿动物 人类大脑疾病的模型现已可用，但神经科学家仍因缺乏技术而受阻。 用于大规模记录和操纵大量经遗传鉴定的神经元的活动 动物实验对象的大脑。Inscope，Inc.是从斯坦福大学剥离出来的，目的是将一种 微型荧光显微镜技术，使神经科学家能够可视化高达1200的钙动态 神经元同时处于清醒状态，在细胞分辨率上表现为啮齿动物。商业银行的发展 系统nVista的部分资金来自之前的NIMH SBIR赠款。这个项目超出了所有人的预期， 举例说明了NIH/NIMH SBIR计划如何在催化创新和支持方面非常有效 神经科学突破性创新的商业化。先前SBIR第一阶段和第二阶段的结果 第二阶段是一个nVista系统，目前在世界各地的200多个实验室使用，这导致了 &gt；在顶级期刊上发表了50篇科学论文。 在此快速通道计划中，我们将在之前成功的SBIR和nVista的成功基础上，开发 并将将nVista转变为多功能平台的下一代大脑地图平台商业化 集成了单色和双色成像、体积成像和光遗传学刺激功能， 以及基于Web、兼容云的数据采集和控制。在第一阶段，我们将设计和制造 用于下一代脑成像平台(AIM 1)的新显微镜原型以及 先进的视频数据采集和处理系统(目标2)。我们将验证这些功能的性能 并与一小群Beta实验室进行第一次活体实验(目标3)。在第二阶段，我们 将制作15个完整的、用户友好的系统，纳入第一阶段(目标4)的反馈，并提供支持 图像处理和分析算法(目标5)。我们将用一个更大的 一套测试版网站，以证明所有功能的科学价值(目标6)。在第二阶段结束时，我们将有一个 新型微型显微镜系统，集成了单色和双色成像、体积成像和 光遗传学刺激能力，以及基于网络的数据采集和控制，为大规模生产做好准备 神经科学界的生产和商业化。这一新产品将取代我们现有的产品 NVista将成为我们的旗舰产品，使世界各地的大脑研究人员能够将神经 脑电活动对大脑功能的影响。我们希望新产品能促进神经回路的研究，并成为 这是世界范围内绘制大脑回路活动图并了解大脑活动的核心工作。我们也 预计该产品将被大型研究中心和Pharma广泛采用，扩大 基础神经科学之外的应用到围绕理解大脑疾病和 开发下一代治疗方法。

Project Summary Our understanding of neuropsychiatric disorders is undergoing a paradigm shift away from simple models emphasizing neurotransmitter imbalance towards more sophisticated theories based on abnormalities in neural circuit function. This is especially important for understanding disorders like Schizophrenia and PTSD, which defy simple biochemical explanations, but are now known to involve abnormal patterns of neural ensemble activity. Even for other types of brain disorders such as neurodegenerative disorders (e.g. Parkinson's Disease), or neurodevelopmental disorders (e.g. Autism) that have molecular-genetic, cellular or environmental causes, there is increasing awareness that the dysfunction is ultimately expressed at the level of neural circuits in the brain, and manifests in abnormal neural circuit dynamics. However, we still lack crucial basic understanding of neural activity patterns during normal behavior and how these patterns are altered in disease. Many rodent models of human brain diseases are now available, but neuroscientists are still hindered by lack of a technology for large-scale recording and manipulation of activity in large populations of genetically identified neurons in the brains of behaving animal subjects. Inscopix, Inc. was spun out of Stanford University to commercialize a miniature fluorescence microscope technology to enable neuroscientists to visualize Ca2+ dynamics in up to 1200 neurons simultaneously in awake, behaving rodents at cellular resolution. The development of the commercial system, nVista, was partly funded by a prior NIMH SBIR grant. The project surpassed all expectations and exemplifies how the NIH/NIMH SBIR program is highly effective in catalyzing innovation and supporting commercialization of breakthrough innovations for Neuroscience. The outcome of the prior SBIR Phase I and Phase II is an nVista system that is today in use at over 200 laboratories around the world and that has led to >50 scientific publications in top-tier journals. In this Fast-Track proposal, we will build on the prior successful SBIR and the success of nVista, and develop and commercialize a next generation brain mapping platform that transforms nVista into a versatile platform integrating single- and dual-color imaging, volumetric imaging, and optogenetics stimulation capabilities, together with web-based, cloud-compatible data acquisition and control. In Phase I, we will design and fabricate prototypes of the new microscope for the next generation brain mapping platform (Aim 1) together with an advanced video data acquisition and processing system (Aim 2). We will validate performance of these prototypes and conduct the first in vivo experiments with a small group of beta labs (Aim 3). In Phase II, we will fabricate 15 complete, user-friendly systems, incorporating feedback from Phase I (Aim 4), with supporting image processing and analysis algorithms (Aim 5). We will perform extensive in vivo experiments with a larger set of beta sites to demonstrate the scientific value of all features (Aim 6). At the end of Phase II, we will have a new miniature microscope system that integrates single- and dual-color imaging, volumetric imaging, and optogenetics stimulation capabilities, together with web-based data acquisition and control ready for mass production and commercialization to the Neuroscience community. The new product will obsolete our current nVista and will become our flagship product enabling brain researchers around the world to causally link neural circuit activity to brain function. We expect the new product to catalyze neural circuit research and to become a centerpiece of worldwide efforts to map brain circuit activity and to understand the brain in action. We also expect the product to be broadly adopted by large research centers and by Pharma, expanding the range of applications beyond basic Neuroscience to translational projects built around understanding brain disease and developing next generation treatments.