ClearScope

结合体内和体外三维(3D)全脑成像的非转基因和 转基因动物模型有望为神经网络连接模式提供新的见解。 对于3D全脑数据集的体外光镜成像，最好的方法是大脑 之后是全脑光片显微镜(LSM)，因为其独特的组合 与共聚焦显微镜和多光子显微镜相比，它具有速度快、3D分辨率高、光毒性低等特点。 与其他方法不同，脑清理/LSM相结合的方法使使用完整组织成为可能 并保留大脑结构内的所有细胞内连接。然而，LSM系统在商业上 不适用于高级3D全脑数据集的体外光镜成像 连接学研究。最近，Raju Tomer博士(Dep.比奥尔。科学，哥伦比亚大学，纽约，NY) 开发了光片theta显微镜(LSTM)，本质上是两个光片的独特排列 倾斜于样品和垂直于样品的一个检测物镜。这部小说 显微镜是LSTM中许多功能的基础，而这些功能并不是任何其他功能都可以使用的 商业上可用的LSM系统。LSTM技术与共焦技术相比具有明显的优势 其他薄片显微镜，包括无与伦比的能力，成像较厚的组织样本在 更大的横向面积(XY)，具有更高的光学分辨率，同时保持快速成像速度，高成像 质量好，光漂白低。这项前景看好的技术是本实验的基础 MarketPlace提议开发ClearScope™，它改进和改进了Tomer博士最初的 LSTM系统创造了一种成功的商业显微镜，供广泛采用。关键技术 将ClearScope开发为商业产品的目标包括创建和测试(I) 基于优化显微镜硬件设计的ClearScope原型；(Ii)新型显微镜 ClearScope的硬件组件，包括一个新的腔体，其中包含被调查的 样品和浸泡介质，以及一种新的检测对象变送器； 用于ClearScope的图像采集软件；重要的是，(Iv)超越 现有最先进的技术将获取的图像堆栈组装成大的3D图像体积 超过10TB，无需对图像信息进行下采样。的生产版本 ClearScope将使神经科学研究界、药理学和生物技术领域受益 R&D，以及整个社会，通过提高对神经网络连接模式的理解 作为大规模连接改变的神经病理基础， 人类的神经精神和神经状况。特别是，这将使基础得到改善。 为复杂的脑部疾病开发新的治疗策略。

Combined in vivo and ex vivo three-dimensional (3D) whole-brain imaging of non-transgenic and transgenic animal models holds the promise of novel insights into neural network connectivity patterns. With regard to ex vivo light microscopic imaging of 3D whole-brain datasets, the best approach is brain clearing followed by whole-brain light sheet microscopy (LSM) because of its unique combination of speed, 3D resolving power, and low phototoxicity compared to confocal and multiphoton microscopy. Unlike other methods, the combined brain clearing / LSM approach makes it possible to use intact tissue and retain all intracellular connections within the brain structure. However, LSM systems commercially available are not suitable for ex vivo light microscopic imaging of 3D whole-brain datasets in advanced connectomics research. Recently, Dr. Raju Tomer (Dept. Biol. Sci., Columbia Univ., New York, NY) developed light sheet theta microscopy (LSTM), essentially a unique arrangement of two light sheets oblique to the specimen and one detection objective perpendicular to the specimen. This novel microscope is the basis for a number of capabilities in LSTM that are not all available with any other commercially available LSM systems. The LSTM technology has distinct advantages over confocal and other light sheet microscopes, including the unmatched ability to image thicker tissue specimens over a larger lateral area (XY) at higher optical resolutions, while maintaining fast imaging speed, high imaging quality, and low photo-bleaching. This promising technology serves as the basis for this Lab to Marketplace proposal to develop the ClearScope™, which refines and improves Dr. Tomer's original LSTM system to create a successful commercial microscope for wide-spread adoption. The key technical objectives for developing the ClearScope as a commercial product include creating and testing (i) a ClearScope prototype based on an optimized microscope hardware design; (ii) novel microscope hardware components for the ClearScope, comprising a novel chamber that contains the investigated specimen and the immersion medium, and a novel detection objective changer; (iii) novel control and image acquisition software for the ClearScope; and importantly, (iv) novel software that surpasses the existing state-of-the-art technology to assemble acquired image stacks into large 3D image volumes exceeding 10TB without need to downsample the image information. The production version of the ClearScope will benefit the neuroscience research community, pharmacological and biotechnological R&D, and society in general by improving understanding of neural network connectivity patterns as well as the neuropathological underpinnings of the large-scale connectional alterations associated with human neuropsychiatric and neurological conditions. In particular, this will result in an improved basis for developing novel treatment strategies for complex brain diseases.