Imaging and Physiology Core

影像和生理学核心

• 批准号: 8059138
• 负责人: RICHARD D MINSHALL
• 金额: $ 23.53万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8059138
• 关键词: Albumins; Area; Blood Vessels; Blood capillaries; Complementary DNA; Confocal Microscopy; Electron Microscopy; Equipment; Filtration; Fluorescence; Fluorescence Microscopy; Fluorescence Resonance Energy Transfer; Gene Delivery; Goals; Human Resources; Image; Image Analysis; Knockout Mice; Life; Liposomes; Lung; Measurement; Methodology; Methods; Modeling; Mus; Participant; Perfusion; Permeability; Photobleaching; Physiological; Physiology; Proteins; Recovery; Research Personnel; Research Support; Resources; Route; Signal Transduction; Specimen; Surface; Techniques; Transfection; Transmission Electron Microscopy; Vascular Permeabilities; capillary; cell fixing; interest; lung vascular injury; monolayer; mouse model; programs; research study; training project

The overall goals of Core D are to provide (i) fiuorescence, confocal, and electron microscopy support, (ii) image analysis, and (iii) physiological support for lung perfusion experiments proposed in all projects. Centralization of the imaging and physiological support within a single core refiects the emphasis that P.l.s have placed on imaging and physiological studies in lung models. Core D is essential in order to fulfill the objectives of all projects. In addition to the research support. Core D personnel will also provide training for project participants in these methodologies. Core D will provide expertise, resources, and equipment for performing lung studies in the knockout mouse models and other mouse models in which proteins of interest are expressed through gene delivery via liposomes. Core D will provide expertise for the transfection of cDNAs in mouse lung microvessels using cationic liposomes. The physiological support component will provide standardized methods for quantification of lung vascular permeability in normal and genetically modified mice. This will include measurement of pulmonary capillary filtration coefficient and vessel wall albumin permeability surface-area product. The methods to be used have been developed specifically for the mouse lung. In addition, lung vascular albumin permeability and the routes of albumin transport will be assessed by electron microscopy and morphometric analysis using described methods. In terms of the imaging component. Core D will provide resources and expertise for (i) live cell and fixed specimen fluorescence, confocal, FRET (fluorescence resonance energy transfer), and TIRF (total internal reflective fluorescence) microscopy and (ii) transmission electron microscopy. Also, Core D will provide assessment of expression of transfected and liposome-delivered proteins by fluorescence and confocal microscopy of endothelial monolayers and whole mount lung sections as described.

核心D的总体目标是提供(I)荧光、共聚焦和电子显微镜支持，(Ii)图像分析，以及(Iii)为所有项目中提出的肺灌注实验提供生理支持。 将成像和生理支持集中在一个核心内，反映了P.L.对肺模型成像和生理研究的重视。核心D对于实现所有项目的目标是必不可少的。除了研究方面的支持。核心D人员还将为项目参与者提供这些方法方面的培训。 Core D将提供专业知识、资源和设备，用于在基因敲除小鼠模型和其他小鼠模型中进行肺部研究，在这些模型中，感兴趣的蛋白质通过脂质体传递基因来表达。CoreD将为利用阳离子脂质体在小鼠肺微血管中导入cDNA提供专业技术。生理支持部分将提供量化的标准化方法 对正常和转基因小鼠肺血管通透性的影响。这将包括测量肺毛细血管滤过系数和血管壁白蛋白通透性表面积乘积。将使用的方法是专门为小鼠肺开发的。此外，肺血管白蛋白 使用所述方法，通过电子显微镜和形态计量学分析来评估渗透性和白蛋白的运输途径。 在成像组件方面。CORE D将为(I)活细胞和固定标本荧光、共焦、FRET(荧光共振能量转移)和TIRF(全内反射荧光)显微镜以及(Ii)透射式电子显微镜提供资源和专业知识。此外，核心D将提供 用荧光和共聚焦显微镜对内皮单分子层和全肺切片的表达进行评估，如所述。