Nanofluidic systems with integrated nanopores for macromolecular sensing, manipulation and confinement

该研究所的研究计划横跨纳米技术/科学和软物质/生物物理领域，致力于发展纳米流体领域。纳米流体器件是芯片上充满流体的通道网络，尺寸约为1-100 nm。这类设备具有分析和操控单分子的能力。例如，纳米通道可以延伸DNA；纳米孔可以通过电子方式检测DNA。这种单分子设备能够在不需要分子放大的情况下进行基因组分析，因此最终可以在单分子、单细胞的基础上进行分析。这种技术将在具有极端细胞间遗传异质性(癌症)和/或可用细胞很少(例如，循环肿瘤细胞的分析)的生物医学系统中得到广泛应用。在实现应用目标的同时，阐明受限大分子的行为将带来新的设备概念和更有效的设计。最后，细胞和病毒本身就是受限的环境。人工的纳米流体结构可以探测宽参数空间的限制，并将活体实验的结果放在全球的物理环境中。在这项提议中，我们将共同从事纳米流体技术的开发和受限聚合物系统的基础物理探索。特别是，我们的重点将是开发一类具有集成纳米孔的新型纳米通道器件。这种纳米孔器件将被用来执行一种新型的横向阻塞传感，它不需要通过孔移位，潜在地导致分辨率不受分子直径的限制，改进的移位控制和高复用能力。这些设备可能导致基于序列特定蛋白质标签和DNA修饰/调节蛋白质的感应的高分辨率DNA电图谱能力。在我们的基础研究轴上，我们将探索DNA在纳米通道中的非平衡动力学和纽结形成，将我们之前的工作扩展到极端限制制度和具有嵌入孔洞的系统。此外，光学镊子和具有集成毛孔的纳米腔将被用于研究在纳米流体“病毒”中的DNA包装，从而导致对推进对噬菌体系统和腔受限聚合物的理解至关重要的定量测量。正如过去HQP的杰出成果所表明的那样，这项工作将带来丰富的多学科培训环境，从而在行业和学术界带来多样化的职业机会。将支持五个HQP(PDF-1、MSC/PHD-3、MSC-1)。HQP将在与聚合物理论家、生物学家、基因组技术和行业专家的合作支持下，在多学科环境中接受纳米制造、单分子技术、单分子数据定量分析、纳米孔传感和聚合物理论方面的培训。

The PI's research program, straddling the fields of nanotechnology/science and soft-matter/biophysics, is devoted to developing the field of nanofluidics. Nanofluidic devices are networks of fluid-filled channels on a chip with dimensions ~1-100 nm. Such devices have the ability to analyze and manipulate single-molecules. For example, nanochannels can extend DNA; nanopores can sense DNA electrically. Such single-molecule devices have the ability to perform genomic analysis without requiring molecular amplification, so that analysis could ultimately be performed on a single molecule, single-cell, basis. Such technology would have wide application in biomedical systems possessing extreme cell-to-cell genetic heterogeneity (cancer) and/or few cells are available (e.g. analysis of circulating tumor cells). Hand in hand with application goals, elucidating the behavior of confined macromolecules leads to new device concepts and more effective design. Finally, cells and viruses are inherently confined environments. Artificial' nanofluidic structures can probe confinement over a wide-parameter space and place results from in vivo experiments in a global, physical context.In this proposal we will jointly pursue nanofluidic technology development and fundamental physics exploration of confined polymer systems. In particular, our focus will be on developing a new class of nanochannel devices with integrated nanopores. The nanochanel-pore devices will be used to perform a novel transverse blockade sensing that does not require through-pore translocation, potentially leading to resolution not limited by the molecule diameter, improved translocation control and high multiplexing capability. These devices could lead to a high-resolution DNA electrical mapping ability based on sensing of sequence specific protein labels and DNA modifying/regulatory proteins. In our fundamental research axis, we will explore DNA non-equilibrium dynamics and knot-formation in nanochannels, extending our previous work to extreme confinement regimes and systems with embedded pores. In addition, optical tweezers and nanocavities with integrated pores will be used to study DNA packaging in a nanofluidic “virus,” leading to quantitative measurements critical for advancing understanding of the phage-system and cavity confined polymers. As demonstrated by past outstanding HQP outcomes, this work will lead to a rich multidisciplinary training environment leading to diverse career opportunities in industry and academia. Five HQP will be supported (PDF-1, MSc/PhD-3, MSc-1). HQP will receive training in nanofabrication, single-molecule techniques, quantitative analysis of single-molecule data, nanopore sensing and polymer theory in a multidisciplinary environment supported by collaborations with polymer theorists, biologists, experts in genomics technology and industry.