AN INTEGRATED PHASE-SPECTRAL FLOW CYTOMETER

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 该研发项目的总体目标是设计、实施、验证和应用一个新的流程 一种将完整荧光发射光谱的收集与 测量和区分荧光寿命。如应用程序简介中所述 在下面的意义部分中，该项目直接在以下几个方面阐述了NIH路线图 通过扩大流式细胞术的筛选和多路扩增能力。我们的 总体方法将是在我们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)细胞计量平台。这将是 通过一系列四个不断发展的目标实现：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- 使用扫描光学编码方法的光谱范围、灵敏度和分辨率。我们会 还集成并进一步扩展了ORCA数字数据采集和控制的能力 以前开发的系统，用于处理来自不断演变的日益复杂的数据 乐器。每个目标都结合了基于珠子和电池的校准和灵敏度的使用 验证不断发展的设计的性能的标准。这样做的好处之一是 不断发展的方法是，我们将在每个阶段都有新的工具可供 协作者，既允许短期应用程序，也允许需要 更高的灵敏度、范围、分辨率和多路复用能力。我们已经建立了四个密钥 在以下领域的合作：蛋白质工程和筛选；开发和应用 新的细胞标记；以及改进的本征光学光谱的检测和测量 用于癌症诊断的组织。我们预计，一旦各种不同的项目 仪器已投入使用，并已移入NFCR的用户设施。其中的几个 这第三个项目的发展也将对中国的另外两个研发项目非常有利 应用，包括提高寿命和光谱测量的范围和分辨率 (项目1)和改进的光谱分辨率和电荷耦合数据处理程序(项目2)。

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).