REAL-TIME SINGLE-MOLECULE CHEMICAL MICROSCOPY

描述（改编自申请人摘要）：本研究项目 旨在开发和应用尖端的分析化学技术， 单分子生物传感器（SMB）和单分子生物传感器（SMEs） 化学显微镜用于实时跟踪一系列生物事件 (e.g., T细胞免疫应答）。 研究 热力学（亲和力、化学计量）和动力学（机理、速率常数） 单细胞表面配体-受体结合反应参数 在SM水平上是了解发病的重要一步 生物级联、疾病感染和潜在药物的设计。 这些创新的方法将证明在解开谜团方面的价值， 目前，我们还无法完全理解癌症等疾病， 和艾滋病从其发病和发展，通过他们的诊断， 治疗 这些研究还将提供独特的机会， 基本理论（例如，锁和钥匙模型）。 最后， 这项研究可能会发现新的细胞信号通路， 生物医学研究新技术的发明。 本发明 以及新型分子器件的开发（例如，SMB）用于生物医学 应用程序也将提出新的挑战，并开辟新的 生物医学工程师的机会。 总体方案已答复 最先进的超灵敏分析和单- 分子检测（SMD）最近描述和证明，在我们的 实验室 两个相互关联的研究目标是：1）开发新的SMB， SM化学显微镜用于单细胞免疫反应的实时跟踪。 将使用SMB与SM的实时耦合来构建仪器 荧光显微镜 这种仪器将能够确定 细胞信号传导的实时热力学和动力学曲线 与单配体-受体相互作用相关的途径 单细胞，以及2）为了在单细胞和SM水平上演示，实时 监测T细胞免疫应答的调节。 SMB将用于 触发单个T细胞表面上的单个配体-受体结合事件 cell. 相关的细胞信号传导途径（例如，水解 肌醇磷脂在质膜，增加细胞内 第二信使水平）将使用SM化学品进行实时监测 显微镜

DESCRIPTION (Adapted from the applicant's abstract): This research project aims to develop and apply cutting-edge analytical chemistry technologies and methodologies such as single-molecule biosensors (SMB) and single-molecule chemical microscopy for real-time tracking of a cascade of biological events (e.g., T cell immune response) in real-time at the SM level. The study of thermodynamic (affinity, stoichiometry) and kinetic (mechanism, rate constant) parameters of ligand-receptor binding reactions on the surface of single cells at the SM level is an essential step in gaining an understanding of the onset of biological cascades, disease infection and the design of potential drugs. These innovative approaches will prove value in unraveling mysteries that presently prohibit us from completely understanding diseases such as cancer and AIDS from their onset and development through their diagnosis and treatment. These studies will also provide unique opportunities to understand fundamental theories (e.g., lock-and-key model) at the SM level. Ultimately, this research may lead to the discovery of new cellular signaling pathways and the invention of novel technologies for biomedical research. The invention and development of novel molecule devices (e.g., SMB) for biomedical applications will also raise the new challenges and open up the new opportunities for biomedical engineers. The general scheme is replied upon the novel approaches of state-of-the-art ultrasensitive analysis and single- molecule detection (SMD) recently described and demonstrated in our laboratory. Two interrelated research aims are: 1) To develop novel SMB and SM chemical microscopy for real-time tracking of single-cell immune response. Instrumentation will be constructed using real-time coupling of SMB with SM fluorescence microscopy. This instrumentation will be capable of determining the real-time thermodynamic and kinetic profiles of cellular signaling pathways associated with single ligand-receptor interaction on the surface of single cells, and 2) To demonstrate at the single-cell and SM level, real-time monitoring of the regulation of T cell immune response. A SMB will be used to trigger a single ligand-receptor binding event on the surface of a single T cell. The associated cellular signaling pathways (e.g., hydrolysis of inositol phospholipids in the plasma membrane, increase in intracellular second messenger levels) will be monitored in real-time using SM chemical microscopy.