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.

化学系的化学测量和成像计划以及国际科学和工程办公室的全球风险基金支持北卡罗来纳大学教堂山分校的Steven A.Sper教授利用这一奖项，对使用基于塑料的柱进行纳米级分离(如电泳)提出了新的见解。活动包括：了解使用压印技术生产塑料纳米柱的技术(用于生产DVD光盘的相同技术)，修改塑料的表面属性，最后，使用纳米尺寸的塑料柱进行具有独特操作特性的分离。更广泛的影响在一定程度上是通过在DNA或RNA测序等许多引人注目的领域中应用纳米级电泳法来展示的，这可以提供由环境影响引起的DNA/RNA分子序列含量变化的信息，识别生物病原体，和/或寻找特定菌株的细菌或病毒感染。该项目团队由美国北卡罗来纳大学和韩国蔚山国家科学技术研究所(UNIST)的研究人员组成，他们建立了富有成效的研究合作。参加这个项目的北卡罗来纳大学的研究生将在UNIST花费12个月的时间来研究这个合作项目的具体目标，利用UNIST的研究基础设施和专业知识，并体验真正的全球研究环境。该项目采用多学科方法，包括转录学、RNA/DNA测序和检测DNA的表观遗传序列变异，以开发使用纳米柱的单分子电泳领域。所使用的柱的不同寻常之处在于，它们包含宽度和深度为纳米尺寸(200 Nm)但长度为50微米的纳米通道。这些通道是使用纳米压印光刻技术在热塑性塑料中制造的。通过监测适当制备的目标的传输来研究小分子和生物聚合物的电泳性；目标被标记有荧光标签，因此可以使用粒子跟踪和荧光显微镜来监测纵向运动的传输动力学，或者使用超分辨率荧光成像来确定横向电迁移。在纳米通道上进行了大量的表面修饰，以了解表面电荷密度的影响以及这些修饰对单分子输运动力学的影响。虽然这些修饰策略已经在微通道中得到了证明，但它们在纳米通道中还没有得到证实。这两种分子的传输都是使用新的超分辨率显微镜进行的，这种显微镜旨在获得前所未有的灵敏度以及空间和时间分辨率。在柱尺寸与德拜长度相似的情况下，该技术还可以帮助阐明电动传输特性。