Bioanalytical and biophysical spectroscopic methods & applications

我的实验室主要研究基于拉曼光谱(RS)和紫外共振拉曼光谱(UVRRS)的生物分析方法和应用。这些是光学非弹性散射技术，可用于在广泛的环境和溶液条件下确定生物分子的浓度和构象。UVRRS提供无与伦比的灵敏度、选择性和背景抑制的组合，研究复杂的生物和/或生化系统。*我们开发了世界上第一个光纤连接的UVRRS系统，用于远程和现场光谱，使UVRRS的力量能够在以前无法访问的环境和应用中得到利用。*这项技术特别适合于使用相对稀薄的微升体积样品来研究其自然状态或封闭/实验操作环境中的生物分子。*该提案从两个相关方向描述了我未来的研究活动：(1)第一个方向是开发基于新的硬件和软件的方法/技术。他的工作包括为UVRRS设计专门的光纤探头和仪器；基于软件的方法包括信号处理和分析工具，以帮助从生物分子中提取和解释包含在拉曼散射信号中的丰富信息。这些方法/技术结合在一起，将使新的应用成为可能，例如，涉及复杂介质中的分析、测量生化动力学、远程(例如，环境)监测、与其他仪器(例如，微流控系统、热循环仪、(2)第二个方向涉及通过新技术实现生物技术应用。我们计划追求的众多应用之一涉及对一些细菌用来降解环境中的酚类化合物(如多氯联苯)的重要酶家族(双加氧酶)成员的进一步结构研究。这项工作的目标之一是使这一家族的酶的工程变种能够用于污染土地和森林的生物修复。

Research in my lab focuses on bioanalytical methods and applications based mainly on Raman spectroscopy (RS) and ultraviolet resonance Raman spectroscopy (UVRRS).  These are optical inelastic scattering techniques that can be used to determine concentrations and conformations of biomolecules under a wide range of environmental & solution conditions.  UVRRS offers an unrivaled combination of sensitivity, selectivity and background rejection needed to investigate complex biological and/or biochemical systems.  We developed the world's first fiber-optic-linked UVRRS system for remote and in situ spectroscopy, enabling the power of UVRRS to be exploited in previously inaccessible environments and applications.  This technology is uniquely suited for investigating biomolecules in their native states or closed/experimentally manipulated environments using relatively dilute, microliter-volume samples.  The proposal describes my future research activities in two related directions:  (1) The first concerns developing new hardware and software based methods/technologies.  Work on 'hardware' technologies includes engineering specialized fiber-optic probes and instrumentation for UVRRS; 'software' based methods include signal processing and analysis tools to aid in extracting and interpreting the rich information contained in the Raman scattering signal from biomolecules.  Together, these methods/technologies will enable new applications involving, for example, analyses in complex media, measuring biochemical dynamics, remote (e.g., environmental) monitoring, experiments in conjunction with other instrumentation (e.g., microfluidic systems, thermal cyclers, calorimeters).  (2) The second direction concerns pursuing biotechnology applications enabled by the new technologies.  One of the many applications that we plan to pursue involves further structural investigations of members of an important family of enzymes (dioxygenases) that some bacteria use to degrade phenolic compounds such as polychlorinated biphenyls (PCBs) in the environment.  One of the goals of this work is to enable the development of engineered variants of enzymes from this family that can be used for bioremediation of polluted lands and forests.