SHARED CONFOCAL MICROSCOPY FACILITY

描述（由申请人提供）：本修订申请要求共聚焦显微镜系统为迈阿密大学医学院神经病学和分子与细胞药理学系以及脑血管疾病研究中心（CVDRC）的NIH资助的研究人员提供服务。描述了NIH资助的六个项目应用领域。在两个程序中，共聚焦显微镜应用于整个动物体内脑缺血模型，以获得血管壁形态，血浆和红细胞流动，白细胞粘附，血栓形成和溶解的高分辨率动态成像。一些程序还利用共聚焦载玻片显微镜进行单标记和多标记免疫组织化学。所要求的系统也将用于共聚焦成像的器官型海马切片制备和活细胞成像。体内应用需要将共聚焦系统放置在我们的动物实验室中，该实验室包含用于显微外科手术、生理监测和基于激光的光血栓形成的仪器和资源。共聚焦仪器将被集成到CVDRC的图像分析核心设施中，开发软件将共聚焦图像映射到多模式图像数据库中，用于多标记免疫组织化学与血流和代谢放射自显影，组织病理学和原位杂交数据集的相关区域分析。NIH资助的六个项目应用领域包括：1）负责光血栓性动脉闭塞诱导的血小板血栓的激光诱导溶解性再通的机制，以及它们与脑中再灌注损伤的相关性。2)局灶性脑缺血：白蛋白神经保护和热疗恶化的机制。3)缺血性脑的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)心肌缺血再灌注中的程序性死亡信号通路及血管新生的治疗作用。

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.