Heterogeneous excited state sorting and analysis cytometry

异质激发态分选和分析细胞术

基本信息

批准号：
7940242
负责人：
Jessica Perea Houston
金额：
$ 31.38万
依托单位：
NEW MEXICO STATE UNIVERSITY LAS CRUCES
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2013-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7940242
关键词：
Address Biological Assay Biology Biomedical Engineering Biomedical Research Cell Cycle Cell Separation Cells Characteristics Chemical Engineering Chemistry Clinical Collaborations Communities Cytometry Data Detection Development Diagnostic Diagnostic Procedure Discipline Energy Transfer Engineering Event Extinction (Psychology)Flow Cytometry Fluorescence Foundations Frequencies Funding Goals Harvest Image Cytometry Imaging Device Individual Investigation Kinetics Laboratories Lead Life Mammalian Cell Measurement Measures Methodology Methods Molecular New Mexico Noise Phase Photons Physics Plague Population Proteins Recovery Relaxation Research Research Training Resolution Sampling Scientist Screening procedure Signal Transduction Sorting - Cell Movement Source Specific qualifier value Spectrum Analysis Stimulus Surface System Techniques Technology Technology Transfer Testing Time Training Uncertainty Universities Validation base bioprocess clinical application cohort digital environmental change fluorophore graduate student heterodyning improved instrument instrumentation intermolecular interaction molecular imaging nanoparticle new technology novel novel diagnostics particle point of care point-of-care diagnostics public health relevance quantum technology development

项目摘要

DESCRIPTION (provided by applicant): This R15 AREA application focuses on the development of a new bioprocess sensing technology poised to enhance biomedical research and training within the New Mexico State University department of chemical engineering. A new diagnostic system is under development for the rapid advancement of temporally-sensitive flow cytometry techniques. That is, new methodologies are proposed for the measurement of heterogeneous excited state fluorescence decays, near- instantaneous inelastic scattering rates, fluorescence lifetime, and phase-filtering approaches from cells and particles in transient states. The proposed time resolved techniques are investigated for real-time cytometric analysis as well as sorting. In addition, these new technologies are developed to identify rare decay kinetic phenomena from individual cells and/or particles, to explore emerging cytometry applications, and for investigation of was to improve and address common cytometry gaps such as autofluorescence noise-plagued assays. To support the development of heterogenous excited-state sorting and analysis cytometry (HESAC), two hypotheses will be examined: (1) that high-throughput multi-exponential fluorescence decay measurements of endogenous cellular emission will not only separate bulk autofluorescence from weak exogenous emissions but also reveal discrete intrinsic species, expose rare events related to intrinsic proteins present at different cell cycles and states, permit separation of Raman scatter for multiplexing, and indicate energy transfer effects; and (2) that the direct selection and screening of stable fluorescent proteins, which are surmised to have distinct heterogeneous lifetimes based on thermally induced conformational changes (i.e. unfolding) and vibrational disruptions (immediate environmental changes), will benefit from fluorescence lifetime-based sorting. The technology development and research questions to be studied in this application have a major potential for widespread influence on the larger cytometry community. Significant contributions to cytometry research are anticipated owing to technological characteristics such as (i) compactness, for point-of-care diagnostics, (ii) multifaceted, for facile commercial integration, (iii) expandable, for development into biomedical molecular imaging devices and systems; and (iv) sophisticated, for the exploration of emerging cytometry assays and never-before studied phenomena. It is projected that the study of high-impact cytometry research will progress into further questions related to sorting, and analysis of cells and particles. Ultimately the most important contribution is that New Mexico State University undergraduate and graduate students be exposed to novel biomedical engineering concepts, methodologies, and principles that coalesce with discoveries in biology, chemistry, and physics disciplines, and that lead to the commercial introduction of quantitative heterogeneous excited-state decay analysis and sorting cytometry. PUBLIC HEALTH RELEVANCE: Development and validation of heterogeneous excited-state-dependent sorting and analysis cytometry (HESAC) for biomedical research applications, clinical diagnostics, and use in point-of-care cytometry systems.

描述（由申请人提供）：该R15区域申请的重点是开发一种新的生物过程传感技术，该技术有望增强新墨西哥州立大学化学工程系内的生物医学研究和培训。正在开发一种新的诊断系统，以快速发展时间敏感的流式细胞仪技术。也就是说，提出了新的方法，用于测量异质激发态荧光衰减，接近瞬时的非弹性散射速率，荧光寿命以及瞬态状态细胞和颗粒的相过滤方法。研究了提出的时间解析技术，以进行实时细胞术分析和分类。此外，这些新技术是为了鉴定单个细胞和/或颗粒的罕见衰减动力学现象，以探索新兴的细胞术应用，并进行研究以改进并解决常见的细胞仪差，例如自荧光噪声噪声质量粘贴的测定。 To support the development of heterogenous excited-state sorting and analysis cytometry (HESAC), two hypotheses will be examined: (1) that high-throughput multi-exponential fluorescence decay measurements of endogenous cellular emission will not only separate bulk autofluorescence from weak exogenous emissions but also reveal discrete intrinsic species, expose rare events related to intrinsic proteins present at different cell cycles and states, permit拉曼散射的分离，用于多重，并指示能量转移效应；（2）基于热诱导的构象变化（即展开）和振动性破坏（即时的环境变化），稳定的荧光蛋白的直接选择和筛选具有明显的异质生命值，将从基于荧光寿命的寿命分类中受益。在本应用程序中要研究的技术开发和研究问题具有对较大的细胞仪社区广泛影响的主要潜力。由于技术特征，例如（i）紧凑性，（ii）多方面，可轻松的商业整合，（iii）可扩展，以开发到生物医学分子成像设备和系统中；（iv）复杂的，用于探索新兴的细胞仪测定法，并且从未研究过现象。预计，对高影响力细胞仪研究的研究将发展为与分类和细胞和颗粒分析有关的进一步问题。最终，最重要的贡献是，新墨西哥州立大学的本科生和研究生将接受新型的生物医学工程概念，方法和原理，这些概念和原理与生物学，化学和物理学学科中的发现结合，并导致定量异型激发型衰减分析和分类细胞综述的商业引入。公共卫生相关性：用于生物医学研究应用，临床诊断和用于护理点的细胞仪系统的异质激发依赖性分类和分析细胞仪（HESAC）的开发和验证。