Single biomolecule sensors using integrated optical waveguides with liquid cores

描述（由申请人提供）：目前，市面上还没有一种生物医学仪器可以提供对单个感兴趣分子的光学灵敏度。单分子研究只能在专门的研究实验室中进行，使用基于各种类型显微镜的昂贵而笨重的设备。在此背景下，本文提出的研究的总体目标和长期目标有两个方面：(1)开发一种新的具有单分子灵敏度的生物医学表征仪器传感器平台；(2)利用这些传感器研究单个生物分子，以更好地了解分子生物学的基本过程。基于集成光波导的新型液芯光波导由两位研究者在NIBIB R21资助下开发。目前，这些波导可以用来引导光通过半导体芯片上的微升体积，并检测几十个分子的荧光，而不需要笨重的外部显微镜。这种新颖的硅基方法与进一步的微流体集成兼容，代表了从微量生物分析物中检测光信号的方式的范式转变。基于这项研究，我们对这个应用程序的具体目标是：单分子灵敏度的演示：我们将改进光波导和检测装置，将灵敏度从目前的40提高到单分子2。集成光电纳米孔波导传感器的开发：利用合成纳米孔作为液核光波导的智能门，我们将开发一种能够同时对单个生物分子进行光电传感的新型传感器。3. 单个核糖体的研究：作为一个具有代表性的生物分子，我们将展示单个荧光标记核糖体的传感，并使用集成波导在毫秒时间尺度上研究动态效应，最终目标是阐明RNA翻译过程的动力学。基于集成技术的单分子水平灵敏度仪器将是紧凑、廉价和便携的。它将对生物学、医学和公共卫生产生重大影响。在世界各地的临床环境、医生办公室、偏远地区和不发达国家广泛部署改进的分析仪器是可能的。此外，分子生物学前沿的研究人员可以关注实验结果，而不是测量仪器。因此，可以获得对药物开发，疾病检测和治疗具有长期影响的基础生命科学的新见解。

DESCRIPTION (provided by applicant): At this time, no commercially available biomedical instrument exists that provides optical sensitivity to a single molecule of interest. Single-molecule studies are only carried out in specialized research labs using costly and bulky setups based on various types of microscopy. In this context, the overall goals and long- term objectives of the research proposed here are two-fold: (I) Development of a new kind of sensor platform with single molecule sensitivity for biomedical characterization instruments, (ii) Use of these sensors to study individual biomolecules to gain better understanding of fundamental processes in molecular biology. Integrated optical waveguides based novel liquid-core optical waveguides were developed by two of the investigators under an NIBIB R21 grant. At present, these waveguides can be used to guide light through Pico liter volumes on a semiconductor chip, and to detect fluorescence from tens of molecules without the need for a bulky, external microscope. This novel silicon-based approach is compatible with further micro fluidic integration and represents a paradigm shift in the way optical signals from minute amounts of biological analytes can be detected. Building on this research, our specific aims for this application are: 1. Demonstration of single-molecule sensitivity: We will improve the optical waveguides and detection setup to improve the sensitivity from currently 40 to single molecules 2. Development of integrated electrical and optical nanopore waveguide sensor: Using synthetic nanopores as smart gates to the liquid-core optical waveguides, we will develop a novel sensor that enables simultaneous electrical and optical sensing of single biomolecules. 3. Study of individual ribosome's: As one representative biomolecule, we will demonstrate sensing of single fluorescently labeled ribosomes and use the integrated waveguides to study dynamic effects on a millisecond timescale with the ultimate goal of elucidating the dynamics of the RNA translation process. An instrument with single molecule level sensitivity based on integrated technology would be compact, inexpensive, and portable. It would have significant impact on biology, medicine, and public health. Widespread deployment of improved analytical instrumentation in clinical settings, doctor's offices, remote locations, and underdeveloped countries around the world would be possible. In addition, researchers at the forefront of molecular biology could focus on the experimental results instead of the measurement apparatus. As a result, new insight into fundamental life science with long-term effects on drug development, disease detection and treatment can be gained.