Metallic Surfaces and Particles in DNA Analysis

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。在这个项目最初的资助期间，我们证明了在非均相银粒子附近的染料标记的dma显示出更高的量子产率，增加的福斯特共振能量转移（FRET）距离和光稳定性。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，我们称之为表面等离子体耦合发射（SPCE）。在下一阶段，我们将扩展这些研究，以使用为特定荧光团和应用而设计的精确纳米制造金属结构。通过将金属结构中的地面和/或激发荧光团与表面等离子体激元耦合，可以获得所需的效果。我们提出以下具体目标：具体目标1：开发用于优化MEF的规则形状粒子阵列，以提高与表面结合的染料标记核酸的检测灵敏度。A.制作各种尺寸、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。B.用单分子光谱法测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，用于结合荧光团标记DNA的PCF表面。制造纳米孔阵列、纳米片和光栅，以获得最佳的荧光团-等离子体耦合。B.开发波长选择性等离子体结构用于DNA分析的定向发射。具体目标3：具体目标1至2中描述的金属结构的应用，以增强当前的DNA阵列平台。A.开发金属纳米结构的DNA杂交方案。B.应用纳米结构阵列识别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）。

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).