基于新型纳米孔阵列-消逝波激发模式的活细胞单分子检测研究

项目名称： 基于新型纳米孔阵列-消逝波激发模式的活细胞单分子检测研究

项目编号： No.21475087

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 高分子科学

项目作者： 董朝青

作者单位： 上海交通大学

项目金额： 87万元

中文摘要： 本项目旨在以贵金属纳米粒子为结构单元，基于纳米粒子表面分子相互作用，组装具有纳米孔阵列结构基质。基于贵金属纳米孔荧光增强效应，建立一种具有高时空分辨的，消逝波激发场的荧光相关光谱单分子探测新方法。研究贵金属纳米粒子具有局域表面等离子体共振（LSPR）荧光增强效应，探究贵金属纳米孔结构增强荧光（NEF）新机理。采用纳米孔结构和全内反射产生消逝波的光学构型显著减小单分子探测方法的检测体积，提高信背（噪）比和单分子探测时空分辨率。探讨纳米孔中分子在消逝波能量场作用下的扩散动力学及理论模型。构建活细胞中荧光相关光谱单分子探测新平台，研究细胞膜的精细结构如膜蛋白在细胞膜上扩散行为和分布等，研究配体与细胞膜上受体相互作用等。

中文关键词： 单分子检测；单细胞分析；纳米孔；消逝波

英文摘要： In this project, new matrix composed of nanopore array structure is assembled based on the interaction of surface molecules in the nanoparticles using noble metal nanoparticles as building units. Based on these noble metal nanopore-enhanced fluorescence (NEF) effect and the introduction of evanescent wave as excitation mode, a novel, highly spatial and temporal fluorescence correlation spectroscopy (FCS)-single molecule detection method is developed for single cell analysis. Local surface plasmon resonance (LSPR) enhanced fluorescence effect of noble metal nanoparticles and the nanopore-enhanced fluorescence (NEF) mechanism is investigated. The novel optical configuration induced by nanopore structure and evanescent wave excitation mode can remarkably reduce the detection volume, improve the ratio of signal to background noise and enhance the spatial and temporal resolution. The diffusion behaviors of biomolecules within nanopore excited in the evanescent wave field are investigated and its mathematical model is simulated. The novel FCS-single molecule detection platform for single living cells is constructed and the fine structure of cell membrane is investigated such as dynamic behavior and distribution of the membrane proteins in the cell membrane. And the interaction of ligand with the receptors in the cell membrane is studied.

英文关键词： single molecule detection;single cell analysis;nanopore;evanescent wave