Metallic Surfaces and Particles in DNA Analysis

DNA 分析中的金属表面和颗粒

• 批准号: 7470691
• 负责人: Joseph R. LAKOWICZ
• 金额: $ 39.24万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-27 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7470691
• 关键词: Biohazardous Substance; Biological Assay; Class; Coupling; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; DNA; DNA Binding; DNA analysis; Detection; Development; Dyes; Energy Transfer; Film; Fluorescence; Funding; Genes; Goals; Gold; Label; Measurement; Metals; Nanostructures; Nucleic Acid Binding; Numbers; Oligonucleotide Probes; Pathogen detection; Photons; Polymorphism Analysis; Primer Extension; Protocols documentation; Shapes; Silver; Spectrum Analysis; Standards of Weights and Measures; Structure; Surface; Testing; base; design; desire; dideoxynucleotide; fluorophore; particle; pathogen; quantum; single molecule; size

DESCRIPTION (provided by applicant): We propose to continue our studies of the use of metallic surfaces and particles for applications in fluorescence based DMA analysis. During the initial funding period of this project we demonstrated that dye- labeled DMAs in the proximity of hetrogeneous silver particles display increased quantum yields, increased Forster resonance energy transfer (FRET) distances and increased photostability. We also showed that fluorophores immobilized near continuous metal surfaces can induce highly efficient directional emission into a substrate, a phenomenon we call surface plasmon-coupled emission (SPCE). In the next period we will extend these studies to use precisely nanofabricated metallic structures designed for use with specific fluorophores and applications. The desired effects will be obtained by coupling of the ground and/or excited fluorophores with surface plasmons in the metal structures. We propose the following specific aims: Specific Aim 1: Development of regular arrays of defined shape particles for optimized MEF to increase the detection sensitivity of dye-labeled nucleic acids bound to surfaces. A. Fabricate metal particles arrays with various sizes, shapes, and spacing, with the goal of maximum sensitivity from different wavelength fluorophores. B. Perform single molecule spectroscopy to determine the photostability of fluorophores near the metallic particles. Specific Aim 2: Development of semi-continuous periodic metallic surfaces for PCF of fluorophore-labeled DNA bound to surfaces. A. Fabricate nanohole arrays, nanorings, and gratings for optimal fluorophore-plasmon coupling. B. Develop wavelength-selective plasmonic structures for directional emission for DNA analysis. Specific Aim 3: Applications of the metallic structures described in Specific Aims 1 to 2 to enhance current DNA array platforms. A. Develop DNA hybridization protocols on the metallic nanostructures. B. Apply the nanostructures array for identification of class B biohazard agents. C and D. Apply the nanostructures for analysis of the cystic fibrosis transmembrane regulator gene (CFTR).

描述(由申请人提供)：我们建议继续研究金属表面和颗粒在基于荧光的DMA分析中的应用。在这个项目的初始资金期间，我们证明了在非均相银颗粒附近的染料标记的DMA表现出更高的量子产率、更长的Forster共振能量转移(FRET)距离和更高的光稳定性。我们还发现，固定在连续金属表面附近的荧光团可以诱导高效的定向发射到衬底中，这种现象我们称之为表面等离子体耦合发射(SPCE)。在接下来的一段时间里，我们将扩展这些研究，以精确地使用纳米级软化金属结构，这些结构设计用于特定的荧光团和应用。通过将地面和/或激发的荧光团与金属结构中的表面等离子体耦合，将获得所需的效果。我们提出了以下具体目标：具体目标1：开发形状规则的粒子阵列，以优化MEF，以提高结合在表面上的染料标记核酸的检测灵敏度。制造不同大小、形状和间距的金属颗粒阵列，目标是从不同波长的荧光团获得最大的灵敏度。进行单分子光谱分析，以确定金属颗粒附近荧光团的光稳定性。具体目标2：开发半连续周期性金属表面，用于与表面结合的荧光标记DNA的PCF。A.制造纳米孔阵列、纳米环和栅格，以实现最佳的荧光团-等离子体耦合。发展用于DNA分析的定向发射的波长选择性等离子体结构。具体目标3：具体目标1至2中描述的金属结构的应用，以增强目前的DNA阵列平台。A.开发金属纳米结构上的DNA杂交方案。B.应用纳米结构阵列鉴定B类生物危害剂。C和D应用纳米结构分析囊性纤维化跨膜调节基因(CFTR)。