Integrated perfusion and confocal imaging system

集成灌注和共焦成像系统

• 批准号: 9075584
• 负责人: Ora A Weisz
• 金额: $ 47.86万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9075584
• 关键词: Animal Model; Bladder; Blood Vessels; Cell physiology; Cells; Cellular biology; Disease; Feedback; Fluorescence Resonance Energy Transfer; Functional disorder; Funding; Housing; Image; Institution; Interdisciplinary Study; Kidney; Life; Membrane Protein Traffic; Microscope; Nephrons; Normal Cell; Perfusion; Pharmacology; Photons; Physiological; Physiology; Process; Proteins; Regulation; Renal tubule structure; Research; Research Personnel; Resolution; Services; Signal Pathway; Signal Transduction; Speed; System; Techniques; Time; Tissues; United States National Institutes of Health; Universities; base; detector; imaging system; intravital microscopy; kidney imaging; knock-down; medical schools; metabolomics; micromanipulator; ranpirnase; tool

 DESCRIPTION (provided by applicant): Microperfusion of dissected tubules is an established technique that can provide information about cellular function within isolated nephron segments in the absence of confounding feedback variables. This information cannot be obtained using other approaches, including intravital microscopy. Recent advances in transduction approaches to express heterologous constructs as well as to efficiently knock down endogenous proteins in kidney and bladder have expanded the opportunity to use live cell approaches, including FRET, to investigate physiologic regulation of membrane trafficking and signaling pathways in real time in isolated tubules. This request is for funds to purchase a perfusion imaging system to enable high speed, high resolution imaging of microperfused kidney tubules and blood vessels as well as other tissues and cells. The system consists of a single photon Nikon A1R confocal microscope with a resonant scanner, spectral detector and FRET module, dual micromanipulators, and an environmental chamber. The system will be housed within the P30-funded George M. O'Brien Pittsburgh Center for Kidney Research at the University of Pittsburgh School of Medicine. The mandate of this center is to facilitate multidisciplinary research, trainin and information transfer related to kidney physiology, cell biology, pharmacology, and pathophysiology. The Center includes cores for Kidney Imaging, Cellular Physiology, Single Nephron and Metabolomics, and Model Organisms, and serves a large group of NIH-funded investigators at the University of Pittsburgh and at other institutions. Our center lacks a dedicated microscope for live imaging of perfused tubules, and the availability of the proposed system will significantly enhance the services we are able to provide to center users. Relevance- The proposed system will provide a crucial tool for the research of numerous biomedical investigators studying the cellular basis of disease. In particular, studies of isolated tubules and vessels conducted with this system are uniquely powerful for elucidating signal transduction processes associated with normal cell function and disease processes.

 描述（由适用提供）：在没有混杂的反馈变量的情况下，可以在隔离的肾单位段内提供有关蜂窝函数的既定技术。无法使用其他方法（包括浸润显微镜）获得此信息。表达异源结构的转导方法的最新进展以及有效地敲低肾脏和膀胱中的内源性蛋白质，扩大了使用包括FRET在内的活细胞方法的机会，以研究隔离管中实时实时的膜运输和信号通路的生理调节。该要求是要购买一种灌注成像系统，以实现对微填充肾管和血管以及其他组织和细胞的高速，高分辨率成像。该系统由一个单个光子Nikon A1R共聚焦显微镜组成，带有顾问扫描仪，光谱探测器和FRET模块，双微型手持器和环境室。该系统将安置在匹兹堡大学医学院的P30资助的George M. O'Brien Pittsburgh肾脏研究中心。该中心的任务是托管与肾脏生理学，细胞生物学，药理学和病理生理学有关的多学科研究，TRARIN和信息转移。该中心包括用于肾脏成像，细胞生理学，单个肾单位和代谢组学以及模型生物体的核心，并为匹兹堡大学和其他机构的大量NIH资助的研究人员提供服务。我们的中心缺乏用于灌注小管的实时成像的专用显微镜，并且所提出的系统的可用性将大大增强我们能够为中心用户提供的服务。相关性 - 拟议的系统将为研究疾病细胞基础的众多生物医学研究人员的研究提供至关重要的工具。特别是，孤立的研究 使用该系统进行的小管和血管对于阐明与正常细胞功能和疾病过程相关的信号转导过程具有独特的功能。