Imaging Cryomicrotome for Whole-Animal Fluorescence Imaging

描述（由申请人提供）：本提案的目标是获得资金，为分子和基因组成像中心增加成像冷冻切片机，该中心是加州大学戴维斯分校的核心设施，为在动物模型中进行体内成像研究提供基础设施和专业知识。我们建议购买一台巴洛科学成像冷冻切片机（Olympia，WA），包括计算机控制的冷冻切片机（能够将12 x 12 x 24 cm的矩形固体切成40 um厚的切片），荧光成像系统（包括Princeton Instruments ES-3200 CCD照相机、Nikon F-mount透镜、Cermax 300瓦氙弧灯以及滤光器和轮），操作系统的计算机硬件和软件。该系统采用连续的冷冻切片机切片，并在编程的切片数量后，激发组织的块面并记录一个或多个图像;由于块面成像，因此图像自动配准。该系统能够在氙弧灯光谱中使用多达5个波长进行荧光激发，并使用多达5个荧光波长进行成像。软件通过对拍摄的图像进行去卷积来重建图像源的位置。该系统将放置在我们的核心设施中，用于支持和帮助NIH资助的不同领域的研究，例如动脉粥样硬化、癌症、气道疾病、靶向分子成像探针开发以及体内成像技术的验证。该系统将直接支持新的靶向诊断和治疗策略的开发，以及表征和更好地理解人类疾病动物模型中正常和患病组织的分子和生化基础。 公共卫生相关性：分子成像领域寻求发现体内成像特定生物靶点和途径的新方法，最终目标是提供患者特异性和基于分子的诊断信息。此外，分子成像方法正在发展，以直接监测分子靶向治疗，细胞治疗和基因治疗。荧光探针是一种关键资源，因为它们可用于标记纳米颗粒、脂质体、抗体、肽和酶底物，用于动物模型的体内和离体研究。拟议的成像cryomicrotome仪器将集成在我们的中心与现有的2D和3D在体内全身小鼠荧光系统，为NIH资助的研究人员提供一个全面的资源，以研究整个动物中的荧光颗粒，分子和报告蛋白的空间和时间分布，空间分辨率范围从几十微米与该仪器，到几毫米与体内技术。这提供了一个强大的平台，用于表征动物疾病模型，并在临床前模型中非侵入性地确定新诊断和治疗策略的有效性。

DESCRIPTION (provided by applicant): The goal of this proposal is to secure funding to add an Imaging Cryomicrotome to the Center for Molecular and Genomic Imaging, a core facility at UC Davis that provides the infrastructure and expertise to conduct in vivo imaging studies in animal models. We propose to purchase a Barlow Scientific Imaging Cryomicrotome (Olympia, WA) that includes a Computer Controlled Cryomicrotome (able to section a 12 x 12 x 24 cm rectangular solid in 40 um thick sections), Fluorescence Imaging System (including a Princeton Instruments ES-3200 CCD camera, Nikon F-mount lens, Cermax 300 watt xenon arc lamp and optical filters and wheels), Computer Hardware and Software to operate the system. The system takes successive cryomicrotome slices and, after a programmed number of slices, excites the block face of the tissue and records one or more images; since the block face is imaged, the images are automatically registered. The system enables fluorescence excitation with up to 5 wavelengths in the xenon arc lamp spectrum and imaging with up to 5 fluorescence wavelengths. Software reconstructs the location of the image source by deconvolving the images taken. The system will be placed in our core facility and used to support and aid NIH-funded research in diverse areas such as atherosclerosis, cancer, airway disease, targeted molecular imaging probe development, and validation of in vivo imaging technologies. The system will directly support the development of new targeted diagnostic and therapeutic strategies, as well as to characterize and better understand the molecular and biochemical basis of normal and diseased tissue in animal models of human disease. Public Health Relevance: The field of molecular imaging seeks to discover new approaches to imaging specific biologic targets and pathways in vivo, with the ultimate goal of providing patient-specific and molecularly-based diagnostic information. In addition, molecular imaging approaches are being developed to directly monitor molecularlytargeted therapies, cellular therapies and gene therapies. Fluorescent probes are a critical resource, as they can be used to label nanoparticles, liposomes, antibodies, peptides and enzyme substrates for in vivo and ex vivo study in animal models. The proposed imaging cryomicrotome instrument will be integrated in our center with existing 2D and 3D in vivo whole-body mouse fluorescence systems to provide a comprehensive resource for NIH-funded researchers to study the spatial and temporal distribution of fluorescent particles, molecules and reporter proteins in the whole animal at spatial resolutions ranging from tens of microns with this instrument, to a few mm with in vivo techniques. This provides a powerful platform for characterizing animal disease models and non-invasively determining the efficacy of new diagnostic and therapeutic strategies in preclinical models prior to translation to the clinic.