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（由申请人提供）：解剖肾小管的微灌注是一种既定技术，可以在没有混杂反馈变量的情况下提供有关分离肾单位段内细胞功能的信息，这些信息无法使用其他方法（包括活体显微镜检查）获得。表达异源构建体以及有效敲低肾脏和膀胱内源蛋白的转导方法扩大了使用活细胞方法（包括 FRET）来研究膜的生理调节的机会该请求旨在提供资金购买灌注成像系统，以实现微灌注肾小管和血管以及其他组织和细胞的高速、高分辨率成像。单光子尼康 A1R 共焦显微镜，配有共振扫描仪、光谱探测器和 FRET 模块、双显微操作器和环境室。该系统将安装在 P30 资助的 George M. O'Brien 内。匹兹堡大学医学院的匹兹堡肾脏研究中心该中心的任务是促进与肾脏生理学、细胞生物学、药理学和病理生理学相关的多学科研究、培训和信息传递。细胞生理学、单肾单位和代谢组学以及模型生物，并为匹兹堡大学和其他机构的一大批 NIH 资助的研究人员提供服务。我们的中心缺乏用于实时成像的专用显微镜。相关性-该系统将为众多研究疾病细胞基础的生物医学研究人员提供重要的工具。 , 隔离研究 使用该系统进行的肾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。