ClearScope

非转基因和非转基因小鼠的组合体内和离体三维（3D）全脑成像 转基因动物模型有望为神经网络连接模式提供新的见解。 关于3D全脑数据集的离体光学显微镜成像，最好的方法是脑 由于其独特的组合， 与共聚焦和多光子显微镜相比，该显微镜具有更高的速度、3D分辨率和更低的光毒性。 与其他方法不同的是，结合大脑清除/ LSM方法可以使用完整的组织 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 现有技术不适用于先进的3D全脑数据集的离体光学显微镜成像 连接组学研究。最近，Raju Tomer博士（Dept.生物科学，哥伦比亚大学，纽约州纽约） 开发了光片θ显微镜（LSTM），基本上是两个光片的独特排列 倾斜于样品，一个检测物镜垂直于样品。这本小说 显微镜是LSTM中许多功能的基础，这些功能在其他任何显微镜中都不可用。 市售的LSM系统。LSTM技术相对于共焦和 其他光片显微镜，包括无与伦比的能力，图像较厚的组织标本超过 在更高的光学分辨率下具有更大的横向面积（XY），同时保持快速成像速度， 质量和低光漂白。这项有前途的技术是本实验室的基础， 市场建议开发ClearScope™，该产品完善并改进了Tomer博士的原始 LSTM系统将创建一个成功的商业显微镜，供广泛采用。的关键技术 将ClearScope开发为商业产品的目标包括创建和测试（i） 基于优化的显微镜硬件设计的ClearScope原型;（ii）新型显微镜 ClearScope的硬件组件，包括一个新的腔室，其中包含所研究的 样品和浸没介质，以及一种新的检测物镜变换器;（iii）新的控制和 ClearScope的图像采集软件;重要的是，（iv）超越 将采集的图像堆栈组装成大型3D图像体积的现有最先进技术 超过10 TB，而不需要对图像信息进行下采样。量产版 ClearScope将使神经科学研究社区、药理学和生物技术受益 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学基础， 人类神经精神和神经病症。特别是，这将改善基础， 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.