Microanalytical Techniques to Study Single Cell Biochemical Processes

研究单细胞生化过程的微量分析技术

• 批准号: 1411993
• 负责人: Susan Lunte
• 金额: $ 65万
• 依托单位: University of Kansas Center for Research Inc
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1411993&HistoricalAwards=false
• 关键词: Microanalytical; Techniques; Study; Single; Cell

With support of the Chemistry Division of the National Science Foundation, Professor Susan Lunte, in collaboration with Professors Christian Schöneich (University of Kansas), Christopher Culbertson (Kansas State University) and Christian Amatore (Centre National de la Recherche Scientifique, CNRS), will develop new analytical methodologies that can be used to investigate chemical reactions in single biological cells. A central focus of the research will be the development of microfluidic devices for manipulating single cells and conducting measurements of cell chemical reactions using electrical and optical techniques. Microfluidic devices are ultrasmall plumbing networks of channels and valves which can precisely manage small volumes of liquids (one-millionth to a few billionths of a liter). Microfluidics thus enable the study of small systems such as living cells, and also the rapid collection of data. This project is a multidisciplinary/international collaboration between four research groups, each providing unique expertise and facilities. Graduate students will be part of an interdisciplinary team and will acquire training in the fields of chemistry, microfluidics, cell biology, and neurobiology. As part of this project, microfluidics will be integrated into the problem-based learning portion of the instrumental analysis course at the University of Kansas.The proposed methods are based on microchip electrophoresis with electrochemical detection. Microchip electrophoresis permits the separation of multiple species, and electrochemical detection is particularly well-suited for interrogating the redox status of cells. The focus of the proposed studies is on methods for the detection and identification of reactive nitrogen and oxygen species and endogenous antioxidants such as glutathione. A high throughput single cell analysis system with combined electrochemical and fluorescence detection that is capable of monitoring cell redox status as a function of cell phenotype and activation state will also be developed. These new tools will be used to investigate the processes involved in inflammation and neurodegeneration at both the chemical and biological levels.

在美国国家科学基金会化学部的支持下，Susan Lunte 教授将与 Christian Schöneich（堪萨斯大学）、Christopher Culbertson（堪萨斯州立大学）和 Christian Amatore（法国国家科学研究中心）教授合作，开发新的分析方法，可用于研究单个生物细胞中的化学反应。 该研究的中心重点是开发用于操纵单细胞并利用电学和光学技术测量细胞化学反应的微流体装置。 微流体装置是由通道和阀门组成的超小型管道网络，可以精确管理少量液体（百万分之一到十亿分之几升）。 因此，微流体能够研究活细胞等小型系统，并能够快速收集数据。 该项目是四个研究小组之间的多学科/国际合作，每个研究小组都提供独特的专业知识和设施。研究生将成为跨学科团队的一部分，并将接受化学、微流体、细胞生物学和神经生物学领域的培训。作为该项目的一部分，微流体技术将被纳入堪萨斯大学仪器分析课程的基于问题的学习部分。所提出的方法基于微芯片电泳和电化学检测。微芯片电泳可以分离多种物质，电化学检测特别适合询问细胞的氧化还原状态。 拟议研究的重点是检测和识别活性氮和氧以及内源性抗氧化剂（如谷胱甘肽）的方法。 还将开发一种结合电化学和荧光检测的高通量单细胞分析系统，能够根据细胞表型和激活状态监测细胞氧化还原状态。 这些新工具将用于在化学和生物水平上研究炎症和神经变性的过程。