AFM Arrays for Single Molecular Mechanics

免疫系统包括各种各样的细胞，这些细胞通过粘附和粘附相互通信。 信号分子这些分子中的许多都表现为“纳米机器”，可以连接，感知，移动， 驱动，仅举几个它们的功能。要了解这种纳米机器的内部工作原理， 控制它们相互作用的动力学特性的表征。我们用原子力 使用原子力显微镜（AFM）直接机械测量几种免疫系统受体的行为， 单对相互作用分子的水平，其扩展并补充了系综测定 通常用于生物化学和生物物理学。这些实验既费力又费时， 对大量随机数据的要求。为了解决吞吐量瓶颈，我们将 开发具有足够灵敏度，带宽和低噪声的AFM阵列，适用于单个 分子实验这将通过集成几种现有的技术-悬臂阵列， 光电探测器阵列和功能化尖端阵列的新技术-声辐射 压力（阿普）致动器阵列。ARP驱动的AFM阵列-蛋白质芯片-将使大量的 同时进行平行的单分子实验，而不是使用单个AFM进行实验 在很长一段时间内连续发生。阿普驱动技术将在单个AFM中进行测试， 随后以1D和2D AFM阵列的形式，因为它们能够进行单分子实验。 利用这些AFM阵列，我们将表征T细胞受体-配体动力学的力调节 交互.更好地了解这些分子的性质和功能可能会导致 免疫系统功能障碍的诊断和治疗策略的发展。拟议 由于AFM阵列的潜在应用，研究也可能具有重要的商业价值 技术用于药物发现和高速药物筛选。此外，像这样的蛋白质芯片可以 提供将分子识别信号转换为机械、光学和电学信号的手段， 通过天然界面将单个生物分子功能与微机电 系统，这可能对纳米生物技术和生物/化学战产生深远的影响 国防鉴定。

The immune system includes a wide variety of cells that communicate with each other via adhesion and signaling molecules. Many of these molecules behave as 'nanomachines' that connect, sense, move, actuate, to name a few of their functions. To understand the inner workings of such nanomachines requires the characterization of kinetic properties that govern their interactions. We have used atomic force microscopy (AFM) to directly measure mechanistically the behavior of several immune system receptors at the level of single pair of interacting molecules, which extends and complement the ensemble assays commonly used in biochemistry and biophysics. These experiments are laborious and time-consuming due to the requirement for a large ensemble of stochastic data. To address the throughput bottleneck, we will develop arrays of AFM with sufficient sensitivity, bandwidth, and low noise that are suitable for single molecule experiments. This will be done by integrating several existing technologies - cantilever array, photodetector array, and functionalized tip arrays on substrates with a new technology - acoustic radiation pressure (ARP) actuator array. The ARP-driven AFM arrays - protein chips - will enable a large number of parallel single molecular experiments at the same time instead of using a single AFM to conduct them sequentially over a long period of time. The ARP actuation technology will be tested in a single AFM and subsequently in the form of 1D and 2D AFM arraysfor their ability to perform single molecule experiments. Using these AFM arrays, we will characterize the force regulation of kinetics of T cell receptor-ligand interactions. Better understanding of the properties and functions of these molecules may lead to the development of diagnostic and therapeutic strategies for immune system dysfunctions. The proposed research may also have significant commercial value because of the potential application of the AFM array technology to drug discovery and high speed drug screening. Moreover, protein chips such as this may provide means for the transduction of molecular recognition signals into mechanical, optical, and electrical signals through a natural interface to link individual biomolecular functions to microelectromechanical systems, which may have far-reaching implications to nanobiotechnology and biological/chemical warfare identification for national defense.