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的总体目标是为所有项目中提出的肺灌注实验提供（i）荧光、共聚焦和电子显微镜支持，（ii）图像分析和（iii）生理支持。 将成像和生理支持集中在单个核心内反映了P.L.s对肺模型中的成像和生理研究的重视。核心D对于实现所有项目的目标至关重要。除了研究支持。核心D人员还将为项目参与者提供这些方法方面的培训。 核心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，其中感兴趣的蛋白质通过脂质体通过基因递送表达。核心D将提供使用阳离子脂质体在小鼠肺微血管中转染cDNA的专业知识。生理支持部分将提供量化的标准化方法 肺血管通透性的变化。这将包括测量肺毛细血管滤过系数和血管壁白蛋白渗透性表面积乘积。已专门针对小鼠肺开发了待使用的方法。此外，肺血管白蛋白 渗透性和白蛋白转运途径将通过电子显微镜和形态测定分析使用所述方法来评估。 在成像方面。核心D将为（i）活细胞和固定标本荧光、共聚焦、FRET（荧光共振能量转移）和TIRF（全内反射荧光）显微镜和（ii）透射电子显微镜提供资源和专业知识。此外，核心D将提供 如所述通过内皮单层和整体肺切片的荧光和共聚焦显微镜评估转染和脂质体递送的蛋白质的表达。