基于AOD-FLIM/CARS多模光学平台监测单个活细胞内RNA合成过程

细胞生理过程、恶性转化及细胞与药物相互作用过程，均与亚细胞结构的分子组成密切相关，活细胞内分子成分始终处动态变化过程中。但目前尚缺乏合适技术用于对活细胞中生物大分子进行动态探测和全面表征，从而局限了实时监测药物-细胞相互作用过程的动态变化。本项目将建立双光子激发荧光成像、动态荧光寿命成像与定量共焦拉曼显微光谱技术相结合的光学方法研究平台，用于一种可定量表征和监控细胞核特定结构功能区域RNA合成过程中的大分子环境，对细胞核内大分子进行多模态光学成像方法并且对相应区域内蛋白质、RNA和DNA等分子分布及浓度进行监测。经过对不同光学方法获得的数据进行整合，实现对肿瘤细胞RNA合成过程中细胞核内相应区域的分子结构和微环境变化进行动态监测，并用于筛选和研究靶向RNA聚合酶的药物。本项目在研究癌细胞RNA合成等生理过程以及抗肿瘤药物筛选等领域，具有重要的研究意义和应用价值。

Fundamental cell biology postulates that the cellular physiologic processes that responds to drug action or malignant transformations tightly correlates with the composition of the subcellular structures. Cellular molecular structure is dynamic and can be rapidly changing. However, there is a lack of established techniques for dynamic analysis and global characterization of molecular organization in live cells, which has limited dynamic monitoring of the interactions between drug and cells. In this proposal we will build a multimodal optical platform encompassing Fluorescence Lifetime Imaging, Two-photon Excited Fluorescence, multimodal imaging and quantitative confocal Raman microspectroscopy. This multimodal optical platform will quantitatively characterize and monitor the macromolecular environment in major structure-function compartments of the cell nucleus during RNA synthesis process, obtain multimodal imaging of biological molecules and carry out concentration measurements and macromolecular distribution monitoring of proteins, RNA, lipids and DNA in cell nucleus. Then we will integrate the data obtained by different optical methods; achieve dynamic quantitative characterization and monitoring of the macromolecular environment and macromolecular structure in major structure-function compartments of the cell nucleus, screen and study drugs targeting RNA polymerase. The research has important significance and application value on the research of physiological processes of cancer cells, screening anticancer drug and other fields.