AN INTEGRATED PHASE-SPECTRAL FLOW CYTOMETER

集成相位光谱流式细胞仪

• 批准号: 8361770
• 负责人: James P Freyer
• 金额: $ 10.04万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361770
• 关键词: Address; Area; Calibration; Cells; Collaborations; Collection; Complex; Data; Detection; Development; Flow Cytometry; Fluorescence; Funding; Goals; Grant; Information Systems; Label; Measurement; Measures; National Center for Research Resources; Optical Methods; Optics; Performance; Phase; Principal Investigator; Protein Engineering; Research; Research Infrastructure; Resolution; Resources; Screening procedure; Series; Sorting - Cell Movement; Source; Spectrum Analysis; Staging; System; Tissues; United States National Institutes of Health; base; cancer diagnosis; cost; data acquisition; design; digital; improved; instrument; research and development

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The overall goal of this R&D Project is to design, implement, validate and apply a new flow cytometer that integrates collection of complete fluorescence emission spectra with the ability to measure and discriminate fluorescence lifetime. As described in Introduction to the application and in the Significance section below, this project directly addresses the NIH Roadmap in several areas through expanding the screening and multiplexing capabilites of flow cytometry. Our general approach will be construct the integrated instrument based on our existing phase- sensitive (PS) and high-resolution spectral (HRS) cytometery platforms. This will be accomplished through a series of four evolving aims: 1) extend the capabilities of the current instruments and data systems; 2) develop and validate an integrated instrument; 3) extend instrumental capabilities using region-encoding optical methods; and 4) further extend the phase- spectral range, sensitivity and resolution using swept approaches to optical encoding. We will also integrate and further expand the capabilities of the ORCA digital data acquisition and control system developed previously to handle the increasingly complex data coming from the evolving instruments. Each Aim incorporates the use of bead- and cell-based calibration and sensitivity standards for validating the performance of the evolving designs. One of the advantages of this evolving approach is that we will have new instruments available at every stage for use by collaborators, allowing both short-term applications as well as more complex uses requiring increased sensitivity, range, resolution and multiplexing capability. We have established four key collaborations in the areas of: protein engineering and screening; development and application of new cellular labels; and improved detection and measurement of intrinsic optical spectroscopy of tissues for cancer diagnosis. We anticipate many other collaborative projects once the various instruments are operational and moved into the User facility of the NFCR. Several of the developments in this third Project will also be very beneficial for the other two R&D projects in the application, including improved range and resolution of lifetime and spectral measurements (Project 1) and improved spectral resolution and CCD data handling routines (Project 2).

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 该研发项目的总体目标是设计，实施，验证和应用新流程 将完整荧光发射光谱的集合和能力集成的细胞仪 测量和区分荧光寿命。 如应用程序介绍中所述 在下面的“重要性”部分中，该项目直接解决了几个NIH路线图 通过扩展流式细胞仪的筛选和多重毛细管的区域。 我们的 一般方法将根据我们现有阶段构建综合仪器 - 敏感（PS）和高分辨率光谱（HRS）细胞仪平台。 这将是 通过一系列四个不断发展的目标完成：1）扩展当前的功能 仪器和数据系统； 2）开发和验证综合仪器； 3）扩展 使用区域编码光学方法的仪器功能； 4）进一步扩展了相位 光谱范围，灵敏度和分辨率使用扫描方法进行光学编码。 我们将 还集成并进一步扩展了逆期难数字数据采集和控制的功能 以前开发的系统以处理来自不断发展的日益复杂的数据 乐器。 每个目标都包含使用珠和细胞基校准和灵敏度的使用 验证不断发展设计的性能的标准。 其中的优势之一 不断发展的方法是，我们将在每个阶段使用新工具供您使用 合作者，允许短期应用程序以及更复杂的用途 提高灵敏度，范围，分辨率和多路复用能力。 我们已经建立了四个钥匙 蛋白质工程和筛查领域的合作；开发和应用 新的蜂窝标签；并改善了对内在光谱法的检测和测量 用于癌症诊断的组织。 我们期望有许多其他合作项目 仪器是运行的，并移至NFCR的用户设施。 几个 第三个项目的发展也将对其他两个研发项目非常有益 应用，包括改善范围和寿命和光谱测量的分辨率 （项目1）和改进的光谱分辨率和CCD数据处理程序（项目2）。