Light-Sheet Fluorescence Microscope

项目摘要 多细胞生物体的生物学是在多个大小的尺度上协调的，从亚纳米尺度的 分子到宏观尺度，到复杂组织、器官和组织中细胞群体的组织范围内的互连 有机体。使用最先进的光片荧光显微镜(LSFM)和光学清晰度 (组织的清晰度)，目前发育、功能障碍和疾病的动物模型可以用 只有在观察和分析微观分析(单个细胞)时才能获得创新的见解 宏观距离(完整的组织、器官、生物体)。此提案代表来自 美国国立卫生研究院资助的内布拉斯加州大学医学中心(UNMC)及周边研究人员 机构(克雷顿大学、男童镇国家研究医院)获得成像、组织研究资金 加工，数据整合设备，Miltenyi超微II，X-Clarity组织水凝胶 聚合和结算系统以及数据集成工作站目前在内布拉斯加州不可用。这 微尺度到宏观尺度的成像类型对于各种动物研究是至关重要的 由癌症生物学家、神经学家、细胞生物学家、生理学家和发育生物学家在 军委会及其周边机构。在众多基础、转化和临床研究中心和计划中 在UNMC，这项提议将主要服务于三个重点领域：1)癌症(弗雷德和帕梅拉·巴菲特癌症 中心)，2)神经科学(整合和转化神经科学中心)，以及3)临床前药物 开发和发现(多名基础、翻译和临床研究人员与附属机构合作 内布拉斯加州医学医院和诊所)。由Jensen Smith博士领导的拟议组合仪表(19年 生物医学成像经验)，由以下董事组成的咨询委员会提供监督 附属的成像核心，将提供一个强大的机会来支持17个由NIH资助的强大用户基础 研究人员。我们结合了开发新的整体/多模式成像的基本要素 UNMC的管道将结构/解剖数据(MRI、SPECT/CT、超声、LSFM)与功能和 分子数据(活体光学成像、光声学、多光子和高级显微镜)以扩展 并提高正在进行的使用动物模型了解人类疾病的研究的严谨性。除了强大的 在用户支持和需求方面，目前的提案具有强大的机构、机构间和机构内支持。AS 作为动物成像技术多模式连续体的一部分，超微II及其相关设备 将提供各种生物现象的强大视角，从而加速翻译研究 旨在预防和治疗人类疾病，包括神经功能/功能障碍、癌症和 药物开发/发现。

Project Summary The biology of multicellular organisms is coordinated across multiple size scales, from the sub-nanoscale of molecules to the macroscale, to tissue-wide interconnectivity of cell populations in complex tissues, organs, and organisms. Using new, state-of-the-art light-sheet fluorescence microscopy (LSFM) and optical clearing (CLARITY) of tissues, current animal models of development, dysfunction, and disease can be appreciated with innovative insights gained only when microscopic analyses (individual cells) are observed and analyzed over macroscopic distances (intact tissues, organs, organisms). This proposal represents a shared request from various NIH-funded researchers at the University of Nebraska Medical Center (UNMC) and surrounding research institutions (Creighton University, Boys Town National Research Hospital) to obtain funding for imaging, tissue processing, and data integration equipment, the Miltenyi UltraMicroscope II, X-Clarity Tissue Hydrogel Polymerization and Clearing System, and data integration workstation not currently available in Nebraska. This type of micro- to macro-scale imaging is of paramount importance for the diverse array of animal studies conducted by cancer biologists, neuroscientists, cell biologists, physiologists, and developmental biologists at UNMC and surrounding institutions. Of the many basic, translational and clinical research centers and programs at UNMC, this proposal will principally serve three focus areas, 1) Cancer (Fred and Pamela Buffett Cancer Center), 2) Neuroscience (Center for Integrative and Translational Neuroscience), and 3) Preclinical drug development and discovery (multiple basic, translational, and clinical researchers aligned with affiliated Nebraska Medicine Hospital and clinics). The proposed instrument cluster led by Dr. Jensen Smith (19 years of biomedical imaging experience) with oversight provided by an advisory committee consisting of Directors of affiliated imaging cores, will provide a robust opportunity to support the strong user base of 17 NIH-funded researchers. We have combined the foundational elements for developing a novel, holistic/multimodal imaging pipeline at UNMC merging structural/anatomical data (MRI, SPECT/CT, Ultrasound, LSFM) with functional and molecular data (intravital optical imaging, photo-acoustics, multiphoton and advanced microscopy) to expand and elevate the rigor of ongoing studies using animal models to understand human disease. In addition to strong user support and demand, the current proposal has strong institutional, inter- and intra-institutional support. As part of a multimodal continuum of animal imaging technologies, the UltraMicroscope II and associated equipment will provide powerful perspectives of various biological phenomena thereby accelerating translational studies aimed at prevention and treatment of human diseases including, neurological function/dysfunction, cancer, and drug development/discovery.