SHARED CONFOCAL MICROSCOPY FACILITY

描述（由申请人提供）：本修订后的申请要求为迈阿密大学医学院神经病学、分子和细胞药理学以及脑血管疾病研究中心（CVDRC）的一组nih资助的研究人员提供共聚焦显微镜系统。描述了六个美国国立卫生研究院资助的项目应用领域。在两个项目中，共聚焦显微镜应用于活体全动物脑缺血模型，获得血管壁形态、血浆和红细胞流动、白细胞粘附以及血栓形成和溶解的高分辨率动态成像。一些程序也利用共聚焦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准备和活细胞成像。体内应用要求共聚焦系统位于我们的动物实验实验室中，该实验室包含显微外科、生理监测和基于激光的光血栓形成的仪器和资源。共聚焦仪器将集成到CVDRC的图像分析核心设施中，并开发软件将共聚焦图像共同映射到多模态图像数据库中，用于与血流和代谢放射自显像、组织病理学和原位杂交数据集相关的多标签免疫组织化学区域分析。美国国立卫生研究院资助的六个项目应用领域包括：1)光血栓性动脉闭塞引起的激光诱导血小板血栓溶解再通的机制及其与脑再灌注损伤的相关性。2)局灶性脑缺血：白蛋白神经保护和高热加重的机制。3)缺血性脑突触改变与神经可塑性。4)慢性脑血管闭塞的后果。5)缺血预处理与线粒体机制。6)程序性死亡信号通路在心脏缺血再灌注中的作用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.