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一个成像冷冻组，该中心是加州大学戴维斯分校的核心设施，提供基础设施和专业知识，以进行动物模型的体内成像研究。我们建议购买Barlow Scientific Imaging cryomicroome (Olympia， WA)，其中包括计算机控制的cryomicroome（能够在40 um厚的切片中切片12 x 12 x 24 cm的矩形实体），荧光成像系统（包括普林斯顿仪器ES-3200 CCD相机，尼康f卡口镜头，Cermax 300瓦氙气弧灯和光学滤光片和车轮），计算机硬件和软件来操作系统。该系统采用连续的冷冻组切片，并在设定切片数量后，激发组织的块面并记录一个或多个图像；由于对块面进行了成像，因此图像自动配准。该系统可实现氙弧灯光谱中多达5个波长的荧光激发和多达5个荧光波长的成像。软件通过对所拍摄的图像进行反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，如动脉粥样硬化、癌症、气道疾病、靶向分子成像探针的开发和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发展，以及表征和更好地了解人类疾病动物模型中正常和病变组织的分子和生化基础。

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.