Development of an Electroactive Fiber-Optic Chip for Optical/Electrochemical Characterization of Planar-Supported Biological Materials

该奖项支持基于光纤的新型仪器的开发，该仪器将允许对基质支持的生物分子膜进行宽带、高灵敏的光谱/电化学表征，包括膜结合蛋白质及其电活性成分。拟议的仪器基于一种新型的电活性光纤芯片(EA-FOC)，它可以提供：1)表面受限和膜结合的生物相关分子的超灵敏吸收和荧光表征；2)微升细胞体积的蛋白质的光谱/电化学(伏安法、安培法或电位法)表征，并使用10-100飞摩尔的表面受限氧化还原试剂；3)荧光检测离子通过膜组件传输过程中信号分子环境的变化。该仪器将首次使集成光波导和电活性集成光波导(EA-IOW)技术的灵敏度增强与基于光纤的分光计的易用性相结合。计划中的工作扩展了PIS以前的工作，以允许检测膜结合生物分子的结构和活性的变化，并扩大应用范围。该仪器将能够研究结构日益复杂的生物分子膜中的表面受限光学/电化学事件，最终将研究重组为平面支撑的脂类双层的膜蛋白。EA-FOC的最终设计预计将提供一个可与各种商业仪器集成的平台，并将为开发多功能、灵敏和选择性的生物和化学传感器平台提供机会。直接参与该项目的学生将接受新仪器的设计、制造、表征和测试方面的培训，并将在光谱/电化学技术、光纤技术和光学光谱的几个方面获得深入的经验，以及将这些工具应用于生物分子薄膜的研究。

This award supports development of a new, fiber-optic-based instrument which will permit broadband, highly sensitive spectro/electrochemical characterization of substrate-supported biomolecular films, including membrane-bound proteins and their electroactive constituents. The proposed instrumentation is based on a new electroactive fiber-optic chip (EA-FOC) that can provide: 1) ultra-sensitive absorbance and fluorescence characterization of surface-confined and membrane-bound biologically relevant molecules, 2) spectro/electrochemical (voltammetric, amperometric or potentiometric) characterization of proteins at microliter cell volumes and using 10-100 femtomoles of surface-confined redox reagent; 3) fluorescence detection of changes in the environment of signaling molecules that occur during ion transport through membrane assemblies. The instrument will make accessible, for the first time, the sensitivity enhancements of integrated optic waveguide and electroactive integrated optic waveguide (EA-IOW) technologies combined with the ease-of-use of fiber-optic based spectrophotometers. The planned effort extends previous work of the PIs to permit detection of changes in the structure and activity of membrane-bound biomolecules and increase the range of applications. The instrument will be able to study surface-confined optical/electrochemical events in biomolecular films of increasing architectural complexity, culminating with studies of membrane proteins reconstituted into planar supported lipid bilayers. The final design of the EA-FOC is expected to provide a platform that can be integrated with a wide range of commercially available instrumentation, and will provide opportunities for the development of versatile, sensitive, and selective bio- and chemical sensor platform. Students directly involved in the project will be trained in the design, fabrication, characterization, and testing of new instrumentation, and will acquire an in-depth experience in several aspects of spectro/electrochemistry techniques, optical fiber technologies, and optical spectroscopies, and in the application of those tools into studies of biomolecular films.