LUMINESCENCE IMAGING: MOLECULES, CELLS AND TISSUES

发光成像：分子、细胞和组织

• 批准号: 7335017
• 负责人: David Spray
• 金额: $ 20.88万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-15 至 2007-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7335017
• 关键词: astrocytes; astrocytoma; bone marrow; cytokine; electromagnetic radiation; gap junctions; gene expression; genes; housing; light intensity; luminescence; medicine; membrane channels; tissue /cell culture; tissues; training; transplantation; university

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): The purpose of this application is to obtain a photon-counting camera and accessories to allow high temporal and spatial resolution of luminescence within cultured cells and in whole animals; accessories include both multiwell plate and single tube luminometers required for optimization of expression and activation of the photoproteins in vitro and an imaging station in which whole animal imaging will be performed. There is no such high sensitivity camera or whole animal imager currently at Einstein College of Medicine, and there is currently neither type of luminometer in the nine-floor Kennedy Center housing the Department of Neuroscience, where the proposed studies will be performed. The studies that are proposed are all natural extensions of NIH-funded projects of nine investigators that will use the relatively new technique of spatially resolved luminescence recording that has become more accessible due to new generations of vector constructs in which the photoproteins have been optimized with regard to intensity and wavelengths of light output and destabilized with regard to gene expression reporting. The studies of users outlined in this proposal are diverse, including bioluminescent resonance energy transfer to determine dimerization and binding partners for connexin molecules, luminescent reporter molecules to localize ATP release from within astrocytes and astrocytoma cells, luminescence-tagged bone marrow cells in order to characterize their tissue distribution following transplantation into mice, and gene promoters fused to luminescent reporter molecules to identify time course of gene activation in cytokine-stimulated astrocytes and to develop a moderately high throughput assay by which to identify connexin-specific gap junction channel blockers. The proposal also includes the plan for a bimonthly user group meeting, through which new users will be attracted to the instrument and new analytical tools and protocols will be discussed, as well as problems that may arise. It also includes plans for training as well as inclusion of relevant methods in a graduate student c

该子项目是利用 NIH/NCRR 资助的共享仪器补助金提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中出现。列出的机构是资助机构，不一定是研究者的机构。描述（由申请人提供）：本申请的目的是获得光子计数相机和配件，以实现培养细胞和整个动物内发光的高时间和空间分辨率；附件包括优化体外发光蛋白表达和激活所需的多孔板和单管光度计，以及进行整个动物成像的成像站。爱因斯坦医学院目前没有如此高灵敏度的相机或整体动物成像仪，并且神经科学系所在的九层肯尼迪中心目前也没有任何类型的光度计，而拟议的研究将在那里进行。所提出的研究都是 NIH 资助的九名研究人员项目的自然延伸，这些项目将使用相对较新的空间分辨发光记录技术，由于新一代载体结构（其中发光蛋白已在以下方面进行了优化），该技术变得更容易使用光输出的强度和波长以及基因表达报告的稳定性。该提案中概述的用户研究是多种多样的，包括生物发光共振能量转移以确定连接蛋白分子的二聚化和结合伴侣、发光报告分子以定位星形胶质细胞和星形细胞瘤细胞内的 ATP 释放、发光标记的骨髓细胞以表征它们移植到小鼠体内后的组织分布，以及与发光报告分子融合的基因启动子，以识别细胞因子刺激的星形胶质细胞中基因激活的时间过程，并开发一种中等高通量的测定法，通过该测定法识别连接蛋白特异性间隙连接通道阻断剂。该提案还包括计划每两个月举行一次用户组会议，通过该会议将吸引新用户使用该仪器，并讨论新的分析工具和协议以及可能出现的问题。它还包括培训计划以及将相关方法纳入研究生课程中。