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

PI的研究项目横跨纳米技术/科学和软物质/生物物理学领域，致力于发展纳米流体学领域。纳米流控器件是在尺寸为1-100纳米的芯片上由充满流体的通道组成的网络。这种设备具有分析和操纵单分子的能力。例如，纳米通道可以扩展DNA；纳米孔可以电感应DNA。这种单分子设备能够在不需要分子扩增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。这种技术将广泛应用于具有极端细胞间遗传异质性（癌症）和/或可用细胞很少（例如分析循环肿瘤细胞）的生物医学系统。与应用目标携手并进，阐明受限大分子的行为导致新的设备概念和更有效的设计。最后，细胞和病毒是固有的密闭环境。人工纳米流体结构可以探测宽参数空间的约束，并将体内实验的结果置于全局物理环境中。在本课题中，我们将共同致力于纳米流体技术的发展和受限聚合物体系的基础物理探索。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。纳米通道孔器件将用于执行一种新的横向封锁传感，不需要通过孔易位，可能导致不受分子直径限制的分辨率，改进的易位控制和高复用能力。这些设备可以基于序列特异性蛋白质标签和DNA修饰/调节蛋白的传感，实现高分辨率DNA电作图能力。在我们的基础研究中，我们将探索DNA非平衡动力学和纳米通道中的结形成，将我们之前的工作扩展到具有嵌入孔的极端约束制度和系统。此外，光学镊子和集成孔的纳米空腔将用于研究纳米流体“病毒”中的DNA包装，从而导致定量测量，这对于推进对噬菌体系统和腔限制聚合物的理解至关重要。正如过去卓越的HQP成果所证明的那样，这项工作将为工业界和学术界提供丰富的多学科培训环境，从而带来多样化的职业机会。将支持5个HQP （PDF-1, MSc/PhD-3, MSc-1）。HQP将在多学科环境下接受纳米制造、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，并与聚合物理论家、生物学家、基因组学技术专家和工业界合作。

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.