基于上转换发光的新型糖基荧光探针的设计、合成及其在细胞成像中的应用

There are many problems exist in current fault fluorescence detection and imaging technology in the detection of biological tissue, amino acid, hemoglobin and other biological macromolecules and related ions, such as the ultraviolet visible photoluminescence absorption is weak, fluorescence efficiency is low and biological damage area is very large. The project for the existing fluorescent probes in cell membrane permeability difference, the biological utilization degree is low, fluorescence recognition capacity is weak and other issues. Design and synthesis of a class of novel sugar-derived fluorescent probes with conversion luminescence properties. The sugar ring was introduced not only improving the probe membrane permeability and specific absorption, improving the biological utilization degree of conversion, and increaseing coordination and recognition function. The two-photon fluorescence microscopy imaging function of this kind of probes in the living cell and tissue can be achieved for the ratio of detection of small molecular visualization or metal ion. This research work can make function of carbohydrate getting further abundant and development, promoting the structure of living cells and its internal material and having a very important significance for practical real-time dynamic observation and detection.

针对当前断层荧光探测和成像技术在检测生物组织内蛋白质、氨基酸、血红蛋白等生物大分子和其他相关离子时发色团对紫外可见激发光的吸收弱、荧光效率低和光生物损伤区域大等难题，而现有的双光子发光材料在生物组织或细胞中存在着膜透过性差、生物利用程度低、荧光识别能力弱等问题，项目设计了一类结构易于修饰、环境稳定性好、发光效率高且生物相容性好的的新型糖基荧光探针，拟开展该类探针的高效合成方法和细胞成像功能研究，通过在荧光探针上引入糖基来提高探针的膜透过性和特异吸收性，进而提高上转换的生物利用程度，同时可增强糖基上转换荧光探针的配位和识别功能，进一步提高探针的荧光效率。通过本项目研究，可以实现该类探针在细胞内的可视化的比率检测小分子或金属离子的双光子荧光显微成像功能，这将不仅使糖类化合物的功能得到进一步的丰富和拓展，而且对推动活细胞的结构及其内部物质进行实时动态观察和检测的实用化具有十分重要的意义。

具有生物相容性好和荧光效率高的荧光探针是当前断层荧光探测和成像技术发展的关键所在，现有技术在检测生物组织内金属离子和生物大分子时存在发色团吸收弱、荧光效率低和光生物损伤区域大等问题。为此，本项目设计了一类新型糖衍生的荧光探针，开展了负载金属催化制备官能化糖模块和利用新型碳杂键构建杂环砌块的研究，解决了糖基探针制备过程中糖环构型难以调控、官能团修饰复杂、分离提纯困难等难题。对目标探针进行了详细表征，开展了紫外吸收光谱，荧光光谱和金属选择性实验等研究，探索了活细胞内检测金属离子的双光子荧光显微成像性能，新型糖基荧光探针具有结构易于修饰、环境稳定性好、发光效率高且生物相容性好等优点，并提出了探针在金属识别过程中可能的作用机制，为实现荧光探针在细胞内的可视化的金属离子的荧光显微成像功能提供新的方法。