Light-Sheet Fluorescence Microscope

项目摘要 多细胞生物体的生物学是在多个尺寸尺度上协调的，从亚纳米级的 分子到宏观尺度，到复杂组织，器官和组织中细胞群的组织范围内的相互连接， 有机体使用新的、最先进的光片荧光显微镜（LSFM）和光学透明 由于组织的生物学特性，目前的发育、功能障碍和疾病的动物模型可以通过 只有在显微镜下观察和分析（单个细胞）， 宏观距离（完整组织、器官、生物体）。这一提议是来自以下各方的共同请求： 内布拉斯加大学医学中心（UNMC）的各种NIH资助的研究人员和周围的研究 机构（Creighton大学，Boys Town国家研究医院）获得成像，组织 处理和数据集成设备，Miltenyi UltraMicroscope II，X-Clarity组织水凝胶 聚合和清算系统，以及数据集成工作站目前在内布拉斯加州不可用。这 一种从微观到宏观尺度的成像对于各种各样的动物研究是至关重要的 由癌症生物学家、神经科学家、细胞生物学家、生理学家和发育生物学家在 联合国军事委员会和周围机构。在众多的基础、转化和临床研究中心和项目中， 在联合国地雷行动中心，这一建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（综合和转化神经科学中心），和3）临床前药物 开发和发现（多个基础，翻译和临床研究人员与附属机构保持一致） 内布拉斯加州医学医院和诊所）。由詹森史密斯博士（19岁）领导的拟议仪表组 生物医学成像经验），并由以下董事组成的咨询委员会进行监督 附属成像核心，将提供一个强大的机会来支持17个NIH资助的强大用户群 研究人员我们已经结合了开发一种新的整体/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、多光子和先进显微镜），以扩大 并提高正在进行的使用动物模型来了解人类疾病的研究的严谨性。除了强大的 鉴于用户支持和需求，目前的建议得到了机构、机构间和机构内的大力支持。作为 UltraMicroscope II和相关设备是动物成像技术的多模式连续体的一部分 将提供各种生物现象的强大视角，从而加速转化研究 旨在预防和治疗人类疾病，包括神经功能/功能障碍、癌症，以及 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.