SHARED CONFOCAL MICROSCOPY FACILITY

描述(由申请人提供)：本修订申请要求使用共聚焦显微镜系统，为迈阿密大学医学院神经病学、分子和细胞药理学系以及脑血管疾病研究中心(CVDRC)的一组由NIH资助的研究人员服务。介绍了美国国立卫生研究院资助的六个项目的应用领域。在两个项目中，共聚焦显微镜被应用于活体全动物脑缺血模型，以获得高分辨率的动态成像血管壁形态，血浆和红细胞流动，白细胞黏附，以及血栓的发展和溶解。几个项目还利用共焦载玻片显微镜进行单标记和多标记免疫组织化学。所要求的系统还将用于器官型海马片制备的共聚焦成像和Livecell成像。活体应用要求共焦系统位于我们的动物实验实验室内，这些实验室包含用于显微手术、生理监测和基于激光的光血栓形成的仪器和资源。该共焦仪器将集成到CVDRC的图像分析核心设施中，并开发用于将共焦图像共同映射到多模式图像数据库中的软件，用于多标记免疫组织化学与血流和新陈代谢放射自显影、组织病理学和原位杂交数据集的相关区域分析。美国国立卫生研究院资助的6个项目的应用领域包括：1)激光诱导光凝动脉闭塞引起的血小板血栓溶解再通的机制及其与脑再灌注损伤的关系。2)局灶性脑缺血：白蛋白神经保护和高热加重的机制。3)脑缺血后突触改变和神经可塑性。4)慢性脑血管闭塞的后果。5)缺血预适应与线粒体机制。6)心肌缺血再灌流中的程序性死亡信号通路及血管生成的治疗作用。

DESCRIPTION (provided by applicant): This revised application requests a confocal microscopy system to serve a group of NIH-funded investigators in the Departments of Neurology and Molecular and Cellular Pharmacology, and the Cerebral Vascular Disease Research Center (CVDRC), at the University of Miami School of Medicine. Six NIH-funded program application areas are described. In two programs, confocal microscopy is applied to whole-animal brain-ischemia models in vivo to obtain high-resolution dynamic imaging of vascular wall morphology, plasma and red cell flow, leukocyte adherence, and thrombus development and lysis. Several programs also utilize confocal slide-microscopy for single- and multiple-label immunohistochemistry. The requested system will also be used for confocal imaging of the organotypic hippocampal slice preparation and for livecell imaging. The in vivo applications require that the confocal system be located within our animalexperimentation laboratories, which contain instrumentation and resources for microsurgery, physiological monitoring, and laser-based photothrombosis. The confocal instrument will be integrated into the CVDRC's Image-Analysis Core facility, and software developed to co-map confocal images into a multimodal image database for correlative regional analysis of multi-label immunohistochemistry with blood-flow and metabolism autoradiography, histopathology, and in situ hybridization data sets. The six NIH-funded program application areas include: 1) The mechanisms responsible for laser-induced lytic recanalization of platelet thrombi induced by photothrombotic arterial occlusion, and their relevance to reperfusion injury in the brain. 2) Focal cerebral ischemia: mechanisms of albumin-neuroprotection and hyperthermic exacerbation. 3) Synaptic alterations and neuroplasticity in ischemic brain. 4) The consequences of chronic cerebrovascular occlusions. 5) Ischemic preconditioning and mitochondrial mechanisms. 6) Programmed death signaling pathways in cardiac ischemia and reperfusion, and the therapeutic role of angiogenesis.