Single biomolecule sensors using integrated optical waveguides with liquid cores

描述(由申请人提供)：目前，还没有商业上可用的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，使用基于各种类型的显微镜的昂贵和笨重的设备。在此背景下，本研究提出的总体目标和长期目标是双重的：(I)开发一种用于生物医学表征仪器的新型单分子灵敏度传感器平台；(Ii)利用这些传感器来研究单个生物分子，以更好地了解分子生物学的基本过程。其中两名研究人员在NIBIBR21的资助下开发了基于集成光波导的新型液芯光波导。目前，这些波导可以用来引导光穿过半导体芯片上的皮升体积，并在不需要笨重的外部显微镜的情况下检测来自数十个分子的荧光。这种基于硅的新方法与进一步的微流控集成兼容，并代表了从微量生物分析物检测光学信号的方式的范式转变。在这项研究的基础上，我们对这一应用的具体目标是：1.单分子灵敏度的展示：我们将改进光波导和检测装置，将灵敏度从目前的40个提高到单分子2.开发集成的电和光纳米孔波导传感器：利用合成纳米孔作为液芯光波导的智能门，我们将开发一种新型的传感器，能够同时对单个生物分子进行电和光检测。3.单个核糖体的研究：作为一种具有代表性的生物分子，我们将展示单个荧光标记的核糖体的传感，并利用集成波导来研究毫秒级时间尺度上的动态效应，最终目的是阐明RNA翻译过程的动力学。基于集成技术的单分子水平灵敏度的仪器将是紧凑、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将是可能的。此外，处于分子生物学前沿的研究人员可以专注于实验结果，而不是测量仪器。因此，可以获得对基础生命科学的新见解，这些科学对药物开发、疾病检测和治疗具有长期影响。

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.