Imaging Cryomicrotome for Whole-Animal Fluorescence Imaging

描述(申请人提供)：这项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微型机，该中心是加州大学戴维斯分校的核心设施，提供在动物模型中进行活体成像研究的基础设施和专业知识。我们建议购买巴洛科学成像冷冻显微镜(华盛顿州奥林匹亚)，其中包括一台计算机控制的冷冻显微镜(能够将12 x 12 x 24厘米的矩形固体切成40微米厚的部分)、荧光成像系统(包括一台普林斯顿仪器ES-3200 CCD相机、尼康F安装透镜、Cermax 300瓦氙弧灯、光学过滤器和轮子)、计算机硬件和软件来操作该系统。该系统获取连续的冷冻显微切片，并在编程数量的切片之后，激发组织的块面并记录一个或多个图像；由于块面被成像，图像被自动配准。该系统支持氙弧灯光谱中最多5个波长的荧光激发，以及最多5个荧光波长的成像。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，如动脉粥样硬化、癌症、呼吸道疾病、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有针对性的诊断和治疗策略的开发，以及在人类疾病的动物模型中表征和更好地了解正常和疾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新的方法来成像体内特定的生物目标和途径，最终目标是提供患者特定的和基于分子的诊断信息。此外，正在开发分子成像方法来直接监测分子靶向治疗、细胞治疗和基因治疗。荧光探针是一种重要的资源，因为它们可以用来标记纳米颗粒、脂质体、抗体、多肽和酶底物，用于动物模型的体内和体外研究。拟议的成像冷冻微型仪将在我们的中心与现有的2D和3D活体小鼠全身荧光系统集成，为NIH资助的研究人员提供全面的资源，以研究从该仪器的数十微米到体内技术的几毫米的空间分辨率的荧光粒子、分子和报告蛋白在整个动物体内的时空分布。这为确定动物疾病模型的特征和在转化到临床之前在临床前模型中非侵入性地确定新的诊断和治疗策略的有效性提供了一个强大的平台。

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.