Core--Electron spin resonance

在这项资助中提出的项目的一个统一假设是，活性氧（ROS）在介导正常和病理生理血管事件中发挥重要作用。成像核心的目的是为PPG的参与者提供资源和支持，以进行明场和荧光成像以及荧光激活细胞分选（FACS）测量，作为使用以下方法测试这一假设的实验工具：（a）检测和测量来自转基因动物模型的完整血管组织和/或培养细胞内自由基氧和H2 O2的荧光产物;（B） 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的ROS产生细胞;和（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着提高项目负责人的能力，以可视化ROS生产的位置在各种实验或病理条件下，无论是在组织培养和完整的组织，并将这些形态学数据与其他更定量的方法，测量ROS，如电子自旋共振（ESR）光谱。 具体来说，成像核心将：1.提供专业知识，设施和培训，为个人进行免疫细胞化学，免疫组织化学和组织学的血管组织，包括显微镜的使用和计算机辅助分析。 2.协助使用流式细胞术定量测量自由基氧和H2 O2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.