Single Molecule Electrophoresis using Nanochannel Columns Fabricated in Thermoplastics

使用热塑性塑料制造的纳米通道柱进行单分子电泳

• 批准号: 1660002
• 负责人: Steven Soper
• 金额: $ 35.5万
• 依托单位: University of Kansas Center for Research Inc
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-17 至 2019-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1660002&HistoricalAwards=false
• 关键词: Single; Molecule; Electrophoresis; using; Nanochannel

With this award, the Chemical Measurement and Imaging Program of the Division of Chemistry as well as by the Global Ventures Fund of the Office of International Science and Engineering is supporting Professor Steven A. Soper of the University of North Carolina, Chapel Hill to develop new insights into the use of plastic-based columns for nanoscale separations, such as electrophoresis. Activities include: understanding techniques to produce nano-columns in plastics using imprinting (the same technique used to produce DVD disks), modifying the surface properties of plastics and finally, using plastic columns with nanometer dimensions to undertake separations with unique operating characteristics. The broader impacts are demonstrated in part through the application utility of nanoscale electrophoresis in a number of compelling areas such as DNA or RNA sequencing, which can provide information on variations in the sequence content of a DNA/RNA molecule induced by environmental effects, identifying biopathogens, and/or looking for strain-specific bacterial or viral infections. The project team is comprised of researchers at the University of North Carolina (USA) and at Ulsan National Institute of Science and Technology (UNIST) in South Korea, who have established a productive research collaboration. UNC graduate students participating in this project will spend 12 months at UNIST to work on specific aims of this collaborative project, taking advantage of the research infrastructure and expertise at UNIST and experiencing a truly global research environment.This project takes a multi-disciplinary approach that includes transcriptomics, RNA/DNA sequencing, and detection of epigenetic sequence variations in DNA to develop the area of single-molecule electrophoresis using nanometer columns. The columns that are employed are unusual in that they contain nanochannels with nano-dimensions (200 nm) in width and depth, but lengths of 50 µm. These are fabricated in thermoplastics using nanoimprint lithography. The electrophoresis of both small molecules and biopolymers are investigated by monitoring the transport of the appropriately prepared targets; the targets are labeled with fluorogenic tags so that the transport dynamics can be monitored using particle tracking and fluorescence microscopy for longitudinal motions or super-resolution fluorescence imaging for determining transverse electromigration. A host of surface modifications are invoked on the nanochannels to understand the effects of surface charge density and the homogeneity of these modifications on the transport dynamics of single molecules. While these modification strategies have been demonstrated in microchannels, they are yet unproven in nanochannels. The transport of both molecule types are undertaken using new formats of 'super-resolution' microscopy that aims to achieve unprecedented sensitivity, and spatial and time resolution. The technique may also help to elucidate electrokinetic transport properties in the case where column dimensions are similar to the Debye length.

有了这个奖项，化学划分的化学测量和成像计划以及国际科学与工程办公室的全球风险投资基金会支持北卡罗来纳大学教堂教授史蒂文·A·索珀教授，以开发出对纳米级分离的塑料柱的使用新见解。活动包括：了解使用印迹（用于生产DVD磁盘的相同技术）在塑料中生产纳米柱的技术，并修改塑料的表面特性，最后使用具有纳米尺寸的塑料柱进行独特操作特性的分离。在许多引人注目的区域（例如DNA或RNA测序）中，纳米级电泳的应用实用性可以在某种程度上证明了更广泛的影响，这些领域可以提供有关环境效应诱导的DNA/RNA分子序列含量的变化的信息，从而识别生物症和/或寻求特异性细菌或为特异性细菌提供菌株。该项目团队由北卡罗来纳大学（美国）的研究人员和韩国的Ulsan国家科学技术研究所（UNIST）组成，他们建立了产品研究合作。 UNC graduate students participating in this project will spend 12 months at UNIST to work on specific aims of This collaborative project, taking advantage of the research infrastructure and expertise at UNIST and experiencing a truly global research environment.This project takes a multi-disciplinary approach that includes transcriptomics, RNA/DNA sequencing, and detection of epigenetic sequence variations in DNA to develop the area of​​ ​​single-molecule electrophoresis using nanometer列。所使用的列是不寻常的，因为它们含有宽度和深度的纳米二含量（200 nm），但长度为50 µm。这些是在热塑性塑料中使用纳米印刷光刻制成的。通过监测适当准备的靶标的运输来研究小分子和生物聚合物的电泳。靶标用荧光标签标记，以便可以使用颗粒跟踪和荧光显微镜来监测传输动力学，以进行纵向运动或超分辨率荧光成像，以确定横向电气移动。在纳米通道上调用了许多表面修饰，以了解表面电荷密度的影响以及这些修饰对单分子传输动力学的均匀性。尽管这些修改策略已在微通道中证明，但在纳米通道中仍未得到证实。两种分子类型的运输都是使用“超分辨率”显微镜的新格式进行的，旨在实现前所未有的灵敏度，空间和时间分辨率。在柱尺寸与Debye长度相似的情况下，该技术还可以有助于阐明电动传输性能。