Single biomolecule sensors using integrated optical waveguides with liquid cores

目前，还没有商业上可用的生物医学仪器可以为 单个感兴趣的分子。单分子研究仅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置。在此背景下，总体目标和长期目标- 本文提出的研究的长期目标有两个方面：(I)开发一种新型传感器平台 具有单分子灵敏度的生物医学表征仪器，(Ii)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家公司开发 根据NIBIB R21拨款的调查人员。目前，这些波导可以用来引导光通过 半导体芯片上的皮升体积，并检测数十个分子的荧光 需要一个笨重的外部显微镜。这种新的基于硅的方法与进一步的 微流控集成，代表着从微量的光信号 可以检测到生物分析物。在这项研究的基础上，我们对这一应用的具体目标是： 1.单分子灵敏度论证：我们将改进光波导和探测装置 将灵敏度从目前的40个提高到单分子 2.集成电光纳米孔波导传感器的研制：使用合成纳米孔 作为液芯光波导的智能门，我们将开发一种新型的传感器，使其能够 单个生物分子的同时电学和光学传感。 3.单个核糖体的研究：作为一种具有代表性的生物分子，我们将展示单个核糖体的传感 荧光标记的核糖体，并使用集成波导来研究动态效应 毫秒的时间尺度，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的仪器将是紧凑的， 价格低廉，便于携带。它将对生物学、医学和公共卫生产生重大影响。 在临床环境、医生办公室、偏远地区广泛部署改进的分析仪器 世界各地的发展中国家和欠发达国家都有可能这样做。此外，该研究所的研究人员 分子生物学的前沿可以关注实验结果而不是测量 仪器。因此，对基础生命科学对药物开发具有长期影响的新见解， 可以获得疾病的检测和治疗。

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 picoliter volumes on a semiconductor chip, and to detect fluorescencefrom tens of molecules without the need for a bulky, external microscope. This novel silicon-based approach is compatible withfurther microfluidic 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 ribosomes: 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.