AN INTEGRATED PHASE-SPECTRAL FLOW CYTOMETER

集成相位光谱流式细胞仪

• 批准号: 8169406
• 负责人: James P Freyer
• 金额: $ 15.02万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169406
• 关键词: Address; Area; Calibration; Cells; Collaborations; Collection; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Detection; Development; Flow Cytometry; Fluorescence; Funding; Goals; Grant; Information Systems; Institution; Label; Measurement; Measures; Optical Methods; Optics; Performance; Phase; Protein Engineering; Research; Research Personnel; Resolution; Resources; Screening procedure; Series; Source; Spectrum Analysis; Staging; System; Tissues; United States National Institutes of Health; base; cancer diagnosis; 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. 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 Center, which is not necessarily the institution for the investigator. 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).

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。列出的机构为 研究中心，而研究中心不一定是研究者所在的机构。 本研发项目的总体目标是设计、实施、验证和应用新的流程 细胞计数器，其将完整荧光发射光谱的收集与以下能力整合在一起： 测量和鉴别荧光寿命。 如应用程序简介中所述 在下面的意义部分，该项目直接解决了NIH路线图在几个 通过扩大流式细胞术的筛选和多重化能力， 我们 总体思路是在现有阶段的基础上构建综合工具- 灵敏（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来实现：1）扩展当前的能力 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能力;以及4）进一步扩展相位- 光谱范围，灵敏度和分辨率使用扫描方法光学编码。 我们将 此外，还集成并进一步扩展了ORCA数字数据采集和控制系统的能力 系统开发以前处理日益复杂的数据来自不断发展的 仪器. 每个Aim都结合了基于微珠和细胞的校准和灵敏度 验证不断发展的设计的性能的标准。 这样做的好处之一 不断发展的方法是，我们将在每个阶段都有新的工具， 合作者，允许短期应用以及更复杂的用途，需要 提高了灵敏度、范围、分辨率和多路复用能力。 我们已经建立了四个关键 合作领域：蛋白质工程和筛选;开发和应用 新的细胞标记;和改进的检测和测量的内在光学光谱 用于癌症诊断的组织。 我们期待着许多其他合作项目，一旦各种 仪器可运行，并被移入NFCR的用户设施。 的几 这第三个项目的发展也将非常有利于其他两个研发项目， 应用，包括改进寿命和光谱测量的范围和分辨率 改进光谱分辨率和CCD数据处理程序（项目2）。