细胞膜界面反应双模图像检测新方法及系统研究

In this project, a dual mode image detection method and system have been developed to detect the interface reactions of cell membrane. This new system could realize the specific detection by the surface plasmon-coupled directional emission (SPCDE) and the global detection by the surface plasmon resonance (SPR), which provides a new technology for exploring the physiological functions of the cell membrane. SPCDE and SPR dual-mode image detection mechanism and method will be studied. The fluorescence signal and the SPR signal from the same sample could be obtained and be analyzed at the same time. Based on the MEMS technology, microfluidic technology, nano modification technique and so on, the integrated microfluidic system with a dual mode SPR chip will be designed and fabricated, which as the key module is used to fix cells and to stimulate the living cells online. A dual mode image detection system will be designed and prepared based on our previous researches. Both the adherent cells and the suspended cells can be detected by using the new system. The interaction between cell membrane and extracellular matrix will be studied. In short, this project will provide a new method for study the physiological function of cell membrane. It is an important scientific significance for the exploration of the physiological process of the life, the pathogenic mechanism, the therapy mechanism of the drug and so on. Moreover it has the economic value for the detection of clinical medicine, drug screening and detection of food safety and so on.

本项目提出研究面向细胞膜界面反应的双模式图像检测新方法和系统，同时实现表面等离子体耦合定向荧光(SPCDE)的特异性检测和表面等离子体共振(SPR)的全局检测，为探索细胞膜的生理功能提供一种新技术。研究SPCDE与SPR双模图像检测机理和方法，实现定向荧光和SPR的同时检测与同位分析；结合MEMS技术、微流控技术、纳米修饰技术等，研究集成双模式SPR芯片微流控系统关键模块，实现对活细胞的固定与在线刺激；基于已有SPR研究基础，构建双模式图像检测系统；面向贴壁细胞和悬浮细胞，开展细胞膜与细胞外基质的相互作用应用实验，如细胞膜表面受体与配体、促发剂或抑制剂等之间的相互作用。总之，本项目为研究细胞膜的生理功能提供了一种新的方法，对于探索生命体生理过程、疾病致病机理、药物作用机理等具有重要的科学意义，对于临床医学检测、药物筛选、食品安全检测等领域都具有重要的现实价值。

本项目研制出一种定向荧光与表面等离子体共振(SPR)双模图像检测系统样机，实现了实时、动态监测纳米范围内的界面反应过程，获得定向荧光与SPR信息。SPR折射率检测灵敏度达到10-6 RIU；空间分辨率达μm级。研究出一种荧光检测方法，荧光染料标记的蛋白检测范围是160ng/mL-2×104ng/mL。研究出一种简单、低成本的微流控芯片制备方法，模具可以重复使用13次以上。研制出多种适于活细胞检测SPR芯片及其微流控系统，可用于活细胞的双模检测。研究出一种双模图像检测方法，实现细胞实时、动态刺激和检测，同时获得荧光与SPR信号，实现>50个位点的并行检测。本项目成果推动了SPR技术的发展，也为细胞生物学研究提供新思路。