多波长荧光互相关光谱仪（FCCS）研制

Due to having a high detection sensitivity and the advantages of rapid analysis, fluorescence correlation spectroscopy technique has become one of the primary means for the detection of single molecules. It is an ideal analysis tool for single molecule level biomolecules research and revealing the structure and function of macromolecular. In order to better develop the highly sensitive technology for the detection and monitoring of molecular and molecular interactions in living cells, and not subject to any molecular size and weight restrictions, the project is intended to develop: 1)upgrading single wavelength fluorescence autocorrelation spectrometer to multi-wavelength fluorescence cross-correlation spectrometer, realization and optimization of single molecule detection technology with it; 2) connecting multi-wavelength fluorescence cross-correlation spectrometer with microscope for single-cell research, and simplifying instrument design, assembling and building into a portable device; 3)optimization and application of the modified multi-channel fluorescence cross-correlation spectrometer; constructing fluorescence cross-correlation spectroscopy study system for gene expression and messenger RNA distribution in living cells researches; 4) Using the cross-correlation spectroscopy to monitor and track protein interactions, establishing fluorescence cross-correlation spectroscopy detected platform of protein-protein interactions within living cells; 5) the use of fluorescence cross-correlation spectroscopy for molecular diffusion velocity measurements to obtain molecular weight information, build a series of ultra-sensitive, a wide range of live cell fluorescence cross-correlation spectroscopy system.

由于其具有检测灵敏度高及分析速度快等优点,荧光相关光谱技术已经成为单分子检测的主要手段之一，是单分子水平上的生物分子研究，揭示了大分子的结构和功能的一个理想的分析工具。为了更好地拓展该高灵敏技术对于活细胞体内分子及分子相互作用的检测和监控，并不受任何分子体积及重量的限制，本项目拟开发：1）将单波长荧光自相关光谱仪升级为多波长荧光互相关光谱仪，实现并优化单分子检测技术；2）将实现与显微镜的连接以应用于单细胞研究，简化仪器设计，并组装搭建成可携式装置；3）优化并应用所改良的多通道荧光互相关光谱仪，构建活细胞内基因表达和信使核糖核酸的分布荧光互相关光谱研究系统；4）运用互相关光谱技术监测及追踪蛋白质间的相互作用，建立活细胞内的蛋白质-蛋白质相互作用荧光互相关光谱仪检测平台；5）利用荧光互相关光谱技术进行分子扩散速度测量以获取分子量信息，构建系列超灵敏、多元化的活细胞荧光互相关光谱分析系统。

荧光相关光谱技术因具有检测灵敏度高及分析速度快等优点,已经成为单分子检测的主要手段之一，是在单分子水平上进行生物分子研究、揭示大分子结构和功能的理想分析工具。为了更好地拓展该高灵敏技术对于活细胞体内分子及分子相互作用的检测和监控，并不受任何分子体积及重量的限制，本项目以单波长荧光自相关光谱仪为基础，设计并搭建了高灵敏的多波长荧光互相关光谱仪，并将其与显微镜的连接，实现了单分子检测；简化了仪器设计，并组装搭建成可携式装置FCS，FCCS显微镜。该荧光互相关光谱仪有望建成可应用于一系列超灵敏、多元化的活细胞分析的分析检测平台；利用多波长通道标记不同的生物分子，并研究其互相关信号，达到检测并监控其相互作用的目的。该改进大大地扩大了对生物分子相互作用研究的范围，也扩大了荧光相关光谱仪在生物研究中的应用范围。