IDBR: Type A: Development of Plasmofluidic Microscopy (PFM) for High-Sensitivity, High-Throughput Single-Molecule Studies

宾夕法尼亚州立大学被授予开发集成等离子体流体显微镜(PFM)的奖项，该显微镜可以以简单、紧凑、成本效益高、用户友好的形式进行高通量、高灵敏度的单分子研究。由于其前所未有的能力，可以同时实现高灵敏度和高通量，同时使用相对简单、低成本的格式，建议的PFM具有改变单分子研究领域的潜力，并加速生物学、生物化学和医学等多个学科的发现。预计这项技术将有许多不同的应用，并立即产生积极的影响。例如，远离平衡的分子动力学的研究，蛋白质构象变化和折叠等分子性质的表征，或者蛋白质-蛋白质或蛋白质-DNA相互作用的动力学和动力学的研究。拟议的PFM将通过1)录制和分发关于实验程序的视频，2)招募Beta测试实验室，3)在会议上主持演示会议，以及4)与行业合作伙伴合作来传播。女性和代表性不足的少数族裔学生将广泛参与拟议的研究项目。拟议的PFM利用了等离子体(即光和纳米材料之间的相互作用)和微流体所提供的独特功能。它将传统的基于纳米孔径的传感器的灵敏度提高了20-120倍。除了提高灵敏度，建议的PFM技术还提供了更精确的流体控制，最大限度地减少了样品和试剂的消耗，并降低了设备成本。该项目将在微纳尺度上提高对光、流体和分子相互作用的理解，为等离子体流体学的新兴领域做出贡献。建议的基于PFM的T4复制体的单分子研究也将提高对DNA复制过程的理解，这不仅将阐明生物学和生物化学的基本过程，而且将有助于许多疾病的治疗，如癌症和传染病。

An award is made to The Pennsylvania State University to develop integrated plasmofluidic microscopy (PFM) that can conduct high-throughput, high-sensitivity single-molecule studies in a simple, compact, cost-effective, user-friendly format. With its unprecedented ability to achieve high sensitivity and high throughput simultaneously while using a relatively simple, low-cost format, the proposed PFM has the potential to transform the field of single-molecule studies and accelerate discoveries in many disciplines in biology, biochemistry, and medicine. This technology would be expected to have numerous diverse applications with immediate, positive implications. Examples include the study of molecular dynamics far from equilibrium, the characterization of molecular properties such as protein conformational changes and folding, or the investigation of protein-protein or protein-DNA interaction kinetics and dynamics. The proposed PFM will be disseminated through 1) recording and distributing videos on experimental procedures, 2) recruiting beta-testing labs, 3) hosting demo sessions at conferences, and 4) collaborating with industry partners. Female and underrepresented minority students will participate extensively in the proposed research projects. The proposed PFM takes advantage of unique features offered by both plasmonics (i.e., the interaction between light and nanomaterials) and microfluidics. It improves the sensitivity of traditional nanoaperture-based sensors by 20-120 times. In addition to improved sensitivity, the proposed PFM technique provides more precise fluidic control, minimizes consumption of samples and reagents, and reduces equipment costs. This project will improve understanding on the interactions of light, fluid, and molecules at the micro/nano scale, contributing to the emerging field of plasmofluidics. The proposed PFM-based single-molecule studies on the T4 replisome will also improve understanding of the DNA replication process, which will not only elucidate fundamental processes of biology and biochemistry but also shed light on the therapeutics of many diseases, such as cancer and infectious diseases.