Core--Electron spin resonance

这项资助中提出的项目的一个统一假设是，活性氧物种(ROS)在调节正常和病理生理血管事件中发挥着重要作用。成像核心的目的将是为PPG的参与者提供资源和支持，用于作为实验工具的Brightfield和荧光成像和荧光激活细胞分类(FACS)测量，以使用以下方法来验证这一假设：(A)检测和测量来自转基因动物模型的完整血管组织和/或培养细胞中的自由基氧和过氧化氢的荧光产物； 使用细胞特异性荧光探针或细胞特异性抗体的间接免疫荧光鉴定血管组织和培养细胞中产生ROS的细胞；以及(C)动物模型中细胞增殖、细胞肥大和动脉粥样硬化病变大小的检测、测量和组织学分析。成像核心设施的增加将显著提高项目负责人在组织培养和完整组织中在各种实验或病理条件下可视化ROS产生的位置的能力，并将这些形态数据与其他更定量的测量ROS的方法相关联，例如电子自旋共振(ESR)光谱。 具体地说，成像核心将：1.为个人提供进行免疫细胞化学、免疫组织化学和血管组织组织学方面的专业知识、设施和培训，包括显微镜使用和计算机辅助分析。2.辅助用流式细胞术定量测定氧自由基和过氧化氢的产生。

A unifying hypothesis of the projects proposed in this grant is that reactive oxygen species (ROS) play important roles in mediating both normal and pathophysiological vascular events. The purpose of the Imaging Core will be to provide resources and support to participants of the PPG for brightfield and fluorescence imaging and fluorescence activated cell sorting (FACS) measurements as experimental tools to test this hypothesis using the following methods: (a) the detection and measurement of the fluorescent products of free radical oxygen and H2O2 within intact vascular tissues and/or cultured cells from transgenic animal models; (b) the identification of ROS-producing cells in vascular tissue and cultured cells using cell-specific fluorescent probes or indirect immunofluorescence of cell-specific antibodies; and (c) the detection, measurement and histological analysis of cell proliferation, cell hypertrophy, and atherosclerotic lesion size in animal models. The addition of the Imaging Core facility will significantly enhance the ability of project leaders to visualize the locations of ROS production under a variety of experimental or pathological conditions both in tissue culture and in intact tissue and to correlate these morphological data with other more quantitative methods that measure ROS such as electron spin resonance (ESR) spectroscopy. Specifically the Imaging Core will: 1. Provide expertise, facilities and training for individuals in performing immunocytochemistry, immunohistochemistry, and histology of vascular tissues, including microscope usage and computerassisted analyses. 2. Assist with quantitative measurements of free radical oxygen and H2O2 production using flow cytometry.