Shared Instrumentation: Amnis ImageStream System

共享仪器：Amnis ImageStream 系统

• 批准号: 7794152
• 负责人: LESLEYANN HAWTHORN
• 金额: $ 46.44万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-11 至 2011-03-10
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7794152
• 关键词: Advisory Committees; Apoptosis; Cancer Center; Cancer Research Project; Cell physiology; Cells; Cellular Immunology; Cellular Morphology; Classification; Clinical; Core Facility; Data; Detection; Event; Flow Cytometry; Fluorescence; Funding; Funding Applicant; Genomics; Grant; Image; Image Analysis; Individual; Institution; Location; Molecular; Morphologic artifacts; Movement; Nuclear; Nuclear Translocation; Phosphorylation; Population; Proteins; Quantitative Microscopy; Research; Sampling; Signal Transduction; Spottings; System; Technology; Translational Research; United States National Institutes of Health; Visual; Work; high throughput analysis; instrument; instrumentation; new technology; operation; trafficking

DESCRIPTION (provided by applicant): Funding is requested for the purchase of an Amnis ImageStream multispectral imaging flow cytometer which can perform high-throughput analysis of cell populations and cellular processes. This instrument essentially combines the visual power of microscopy, the quantitative ability of image analysis, and the statistical rigor of flow cytometry in a single platform to create exceptional new experimental capabilities. Although destined to become the single analysis platform of choice for cell analysis, this technology has only recently become available such that there are currently still relatively few of these instruments in research institutions. Compared to traditional flow cytometry, the ImageStream also uses data derived from cell morphology which provides a more comprehensive analysis of the sample, while still providing all of the fluorescent information. The feature that is truly enabling, however, is the added ability to identify the intracellular location of the fluorescence signal through cell morphological analysis. The ImageStream also makes it possible to track signaling events or protein movement in a spatial/temporal manner, within or between cells. To determine whether rare events in the data are real or due to artifacts, individual specific signals can be visualized within a specific image using the ImageStream. This also holds true for accuracy in gating strategies. With the introduction of Extended Depth of Field (EDF) technology, the ImageStream can also generate confocal-like images. This technology really enhances the ability to look at FISH spots and co-localization events. As a result of these technological advances, the ImageStream can perform an extensive range of applications including: nuclear translocation, internalization molecular trafficking, molecular co-localization, morphological classification, nuclear foci spot counting and apoptosis detection. In this proposal we present 13 individual NIH funded grants that would directly benefit from access to this novel technology. Specifically the instrument will enhance projects in genomics studies through the application of FISH and detection of phosphorylation. Work in the fields of cellular immunology will benefit considerably from the capacity of the instrument to detect rare cellular subpopulations and cancer research projects which are dependent on the ability to perform co-localization, nuclear translocation studies, apoptosis, intercellular localization and morphological classification will similarly be enhanced. The ImageStream will be incorporated into the Flow cytmoetry core facility in the MCG Cancer Center and its day to day operation will be overseen by the Flow core facility manager. The PI is the director of the MCGCC core facilities for clinical and translational research who will oversee its operation, act as chair of the advisory committee and resolve issues arising with the MCGCC administration.

描述（由申请人提供）：要求购买AMNIS Imagestream多光谱成像流式细胞仪的资金，该流式细胞仪可以对细胞群体和细胞过程进行高通量分析。该仪器本质上结合了显微镜的视觉能力，图像分析的定量能力以及单个平台中流式细胞仪的统计严格，以创建出色的新实验能力。尽管注定要成为细胞分析首选的单个分析平台，但该技术直到最近才获得，因此目前在研究机构中仍然相对较少的这些工具。 与传统的流式细胞仪相比，成像仪还使用了来自细胞形态的数据，该数据提供了对样品的更全面分析，同时仍提供所有荧光信息。然而，真正启用的特征是通过细胞形态分析鉴定荧光信号的细胞内位置的附加能力。成像驱动还可以以空间/时间的方式在细胞之间或之间以空间/时间方式跟踪信号事件或蛋白质运动。为了确定数据中的罕见事件是真实的还是由于伪影而引起的，可以使用成像驱动器在特定图像中可视化单个特定的信号。对于门控策略的准确性也是如此。随着扩展的景深（EDF）技术的引入，成像流也可以产生类似共聚焦的图像。这项技术确实增强了查看鱼类斑点和共定位事件的能力。 由于这些技术的进步，成像传说可以执行广泛的应用，包括：核易位，内部化分子运输，分子共定位，形态学分类，核灶点计数和凋亡检测。在此提案中，我们提出了13个个人NIH资助的赠款，这将直接受益于对这项新技术的访问。特别是该仪器将通过应用鱼类和磷酸化检测来增强基因组学研究的项目。细胞免疫学领域的工作将从仪器的能力中大大受益，以检测罕见的细胞亚群和癌症研究项目，这些项目取决于执行共定位，核易位研究，凋亡，细胞间定位和形态分类的能力得到增强。 成像级将被纳入MCG癌症中心的流式细胞核心设施中，其日常操作将由流程核心设施管理器监督。 PI是MCGCC临床和转化研究核心设施的主任，该设施将监督其运作，担任咨询委员会主席并解决MCGCC政府引起的问题。