NER: Lab on a Tip-Bioconjugate Silica Nanoparticle Probes on Atomic Force Microscope Cantilever Tips

本课题旨在开发一种基于纳米粒子生物偶联化学与原子力显微镜（AFM）相结合的新型纳米尺度生化测量科学仪器。原子力显微镜和纳米粒子化学的结合将纳米技术最广泛使用的工具（原子力显微镜）和纳米科学最有前途的技术之一（纳米粒子化学）结合在一起。在为期一年的NER框架内，该团队（由纳米粒子化学和原子力显微镜的研究人员组成）将展示AFM悬臂的制造和操作，该悬臂具有集成的化学活性纳米粒子尖端，用于探测配体与受体的相互作用。受体驱动的细胞行为是生物系统中一类重要而多样的功能结构。由于受体能够通过配体结合感知环境，并能够通过信号级联将信号传递到细胞内部，因此它们非常适合指导这些过程。本研究的目的是定量测量配体与其生物受体之间的相互作用。目前，这些基本生物成分的研究受到可用的集成成像工具的限制，如放射性标记配体或通过抗体间接检测。这种特殊的测量有望提供迄今为止无法获得的与细胞受体结合的配体的分辨率。预计该技术将为探索受体功能的基本方面、开发药物筛选的新方法和实施生化系统询问的新技术提供新的机会。具体来说，我们将：1。发展原子力显微镜悬臂尖端二氧化硅纳米粒子功能化的方法。设计、合成和表征新型纳米共轭二氧化硅纳米颗粒在悬臂顶端的原位。通过粘附、拍打和侧向力显微镜测量，研究受体-配体相互作用中的力，目的是测量单分子相互作用。

The proposed research aims to develop a new scientific instrument for nanometer scale biochemical measurement based on the integration of nanoparticle bioconjugate chemistry and atomic force microscopy (AFM). The marriage of AFM and nanoparticle chemistry brings together the most widely used tool for nanotechnology (AFM) and one of the most promising techniques for nanoscience (nanoparticle chemistry). Within the one-year NER framework, the team (comprised of researchers in nanoparticle chemistry and atomic force microscopy) will demonstrate the fabrication and operation of AFM cantilevers with integrated, chemically active nanoparticle tips to probe ligand-receptor interactions. Receptor driven cellular behavior represents an important and diverse class of functional structures in biological systems. Receptors are uniquely suited to direct such processes due to their ability to sense the environment through ligand binding and their ability to transmit this signal to the cell interior via signal cascades. The goal of this proposal is quantitatively measure the interactions between ligands and their biological receptors. Currently, the study of these fundamental biological components is limited by available ensemble imaging tools such as radiolabeled ligands or indirect detection via antibodies.This particular measurement holds the promise of providing heretofore unobtainable resolution of ligands binding to cellular receptors. It is anticipated that the developed technique will offer new opportunities to probe fundamental aspects of receptor function, develop novel approaches to drug screening and implement new technologies for the interrogation of biochemical systems. Specifically, we will:1. Develop methods for the functionalization of an Atomic Force Microscope cantilever tip with silica nanoparticles.2. Design, synthesize, and characterize novel nanoconjugate silica nanoparticles in situ on the cantilever tip.3. Investigate the forces in receptor-ligand interactions through adhesion, tapping and lateralforce microscopy measurements, with the goal of measuring single-molecule interactions.