Microanalytical Techniques to Study Single Cell Biochemical Processes

在国家科学基金会化学部的支持下，Susan Lunte教授与Christian Schöneich教授（堪萨斯大学）、Christopher卡伯特森教授（堪萨斯州立大学）和Christian Amatore教授（国家科学研究中心，CNRS）合作，将开发可用于研究单个生物细胞中化学反应的新分析方法。 研究的一个中心焦点将是开发微流体设备，用于操纵单细胞并使用电学和光学技术测量细胞化学反应。 微流体设备是由通道和阀门组成的超小型管道网络，可以精确地管理小体积的液体（百万分之一到几十亿分之一升）。 因此，微流体技术能够研究活细胞等小系统，并快速收集数据。 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，并将获得化学，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，微流体将被整合到堪萨斯大学仪器分析课程的问题为基础的学习部分。微芯片电泳允许多个物种的分离，电化学检测特别适合于询问细胞的氧化还原状态。 拟议研究的重点是检测和鉴定活性氮和氧以及内源性抗氧化剂（如谷胱甘肽）的方法。 还将开发具有组合的电化学和荧光检测的高通量单细胞分析系统，其能够监测作为细胞表型和活化状态的函数的细胞氧化还原状态。 这些新工具将用于在化学和生物学水平上研究炎症和神经变性的过程。

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.