细胞样本中的明显个体差异体现了单细胞分析对于研究细胞功能与行为的重要性，设计开发高灵敏、高选择、高时空分辨的单细胞原位分析技术是当今生物分析化学领域的一大重要课题。细胞的生理活动是一种动态生化过程，而现有的单细胞检测技术多面向细胞在不同阶段的静态检测。针对单细胞分析领域中现有技术方法的不足，依靠申请人在光学显微成像仪器搭建和在单细胞显微成像分析工作中的经验基础，本项目拟围绕二维纳米材料的可调光电性质、金纳米颗粒表面等离子共振耦合增强拉曼散射与距离调制等离子共振耦合光谱红移三个方面的研究展开，建立包括电化学成像、拉曼成像与暗场成像在内的多模态显微成像技术，实现对单细胞外泌生物分子、黄素介导的胞外电子传递行为、单分子蛋白跨膜内吞等生理过程的高精准、高时空分辨的原位动态分析。

Knowledge on the behavior of an individual cell is of critical importance because even genetically identical cells showed large variation. It has become an essential project to design and develop single-cell in-situ analysis techniques with high sensitivity, high selectivity, and high spatiotemporal resolution in the field of analytical chemistry. Aiming at overcoming the shortcomings of present cellular analytical technologies, we plan to make efforts in fabrication of bio-functionalized nanoprobes, development of highly-spatiotemporal microscopies and highly-sensitive in-situ cellular analytical platforms in this proposal. We plan to construct a cellular electrochemical analytical platform based on two-dimension semi-conductor nanomaterials for in-situ analysis of biological processes at single-cell level. We also plan to monitor the Raman spectra of signaling molecules, for example flavin, released by cells based on the combination enhancement of surface plasmon resonance-coupled scattering and the local electromagnetic field in gaps. Furthermore, we plan to design a subnanometer-precision nano-ruler according to the scattering spectral shift induced by the surface plasmon resonance couple effect between plasmonic nanoparticles, which offers a method for real-time tracing of single-molecule membrane protein endocytosis.