IDBR: Development of Heterogeneous Excited State Flow Cytometry Sorting and Analysis

IDBR：异质激发态流式细胞术分选和分析的发展

• 批准号: 0964127
• 负责人: Jessica Houston
• 金额: $ 26.36万
• 依托单位: New Mexico State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-15 至 2014-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0964127&HistoricalAwards=false
• 关键词: IDBR; Development; Heterogeneous; Excited; State

Phase-sensitive flow cytometry is a traditional and minimally disseminated method of capturing the average fluorescence lifetime on an event-by-event basis. That is, early incarnations of frequency-domain cytometry techniques involved a high degree of complexity with cumbersome analog hardware. Owing to the intricate nature of such techniques, the cytomics community has lacked a widespread and robust means of capturing excited-state decays. This project explores new digital methods as a means to introduce multiple lifetimes for additional separation power in cytometry. These include: (1) introduction of multiple-frequency modulation, (2) implementation of innovative approaches for capturing time-resolved signals through non-modulated digital signal processing, and (3) application of fluorescence lifetime parameters to sorting systems so as to separate cells or particles based on decay kinetic measurements. A cytometry system capable of not only average fluorescence lifetime measurements but also multi-exponential decay is appealing for several reasons which include: improving the separation of autofluorescence from exogenous signals, capturing surface enhanced Raman scattering in flow, and quantifying fluorescence protein expression during cell sorting. The development of a multiple lifetime cytometry instrument will significantly impact the cytometry community at large. A temporal method for sorting and analysis may directly affect a wide range of staining and analysis protocols in flow cytometry, improve accuracy of low-density antigen studies, reveal possible rare events that exhibit unique excited state decays, and allow users of all kinds to implement the capability on their commercial instrument. The advancement of excited-state cytometry techniques will also broaden the participation of underrepresented undergraduate and graduate students at New Mexico State University, a Hispanic Serving Institution. Minorities working on the development of the cytometer will be exposed to high-impact bioengineering, chemical engineering, and other multidisciplinary fields needed for the project outcomes. The end goal for all or part of the described cytometry techniques is the integration into the biotechnology industry. All project outcomes can be perused at the Principle Investigator research website: http://che.nmsu.edu/JPH/index.html.

相敏流式细胞术是一种传统的、传​​播最少的方法，用于逐个事件地捕获平均荧光寿命。 也就是说，频域细胞计数技术的早期版本涉及高度复杂性和笨重的模拟硬件。 由于此类技术的复杂性，细胞组学界缺乏捕获激发态衰变的广泛且可靠的方法。 该项目探索新的数字方法，作为引入多种寿命的手段，以提高细胞计数中的分离能力。 其中包括：（1）引入多频调制，（2）实施通过非调制数字信号处理捕获时间分辨信号的创新方法，以及（3）将荧光寿命参数应用于分选系统，以便根据衰变动力学测量来分离细胞或颗粒。细胞计数系统不仅能够测量平均荧光寿命，而且能够测量多指数衰减，其吸引力有几个原因，包括：改善自发荧光与外源信号的分离，捕获流动中的表面增强拉曼散射，以及在细胞分选过程中量化荧光蛋白表达。 多次寿命细胞计数仪器的开发将对整个细胞计数界产生重大影响。 用于排序和分析的时间方法可能会直接影响流式细胞术中的各种染色和分析方案，提高低密度抗原研究的准确性，揭示可能表现出独特的激发态衰减的罕见事件，并允许各种用户在其商业仪器上实现该功能。 激发态细胞计数技术的进步还将扩大新墨西哥州立大学这一西班牙裔服务机构中代表性不足的本科生和研究生的参与。 致力于细胞仪开发的少数​​群体将接触到高影响力的生物工程、化学工程和项目成果所需的其他多学科领域。 上述全部或部分细胞计数技术的最终目标是融入生物技术行业。 所有项目成果均可在主要研究者研究网站上仔细阅读：http://che.nmsu.edu/JPH/index.html。