具有良好发射光谱分辨率的次氯酸双光子比率分子荧光探针的设计、合成及其生物成像研究

Design, synthesis and biological application of two-photon fluorescent probes for imaging is an important interdisciplinary field of chemisty and biology. Two-photon ratiometric fluorescent probes allow the measurement of fluorescence intensities at two emission wavelengths, which should provide a built-in correction for environmental effects. However, most of the small-molecule two-photon ratiometric fluorescent probes that have been reported thus far have shown serious overlap in the emission peaks (poor-resolved emission spectra) before and after interaction with target analytes. These shortcomings can lead to errors in the measurement of fluorescence intensities and ratios. To address the issues associated with the two-photon ratiometric fluorescent probes, based on our previous developed FRET-based one-photon ratiometric fluorescent probe with well-resolved emission peaks and two-photon fluorophores, this project is designed for developing small-molecule two-photon FRET moleculars as probes platform in which the donor and acceptor have well-resolved emission bands, and then further developing a series of two-photon ratiometric fluorescent probes for bioimaging of HOCl in living cells and tissues. The two-photon ratiometric fluorescent probes in this project have a significant characteristics: well-resolved emission spectra before and after interaction with target analytes, which can avoid the errors in the measurement of fluorescence intensities and ratios, which has a good theoretical guidance and practical significance for developing two-photon ratiometric fluorescent probes with well-resolved emission spectra.

双光子小分子荧光探针的设计、合成及生物成像应用是当前化学和生物等交叉学科的一个重要研究方向。比率荧光探针能同时测量两个不同波长处的荧光强度而通过内标校正环境因素的影响。针对目前双光子比率荧光探针与分析物作用前后所面临发射光谱重叠严重（分辨率差）而导致的检测误差问题, 在本研究小组已有的具有良好发射光谱分辨率的单光子比率荧光探针和双光子荧光染料的研究基础上，本项目旨在发展能量供/受体之间具有良好发射光谱分辨率的双光子荧光共振能量传递分子作为新型荧光探针平台，并基于该平台创建次氯酸双光子比率荧光探针，从而利用双光子比率成像检测组织与细胞内的次氯酸。本项目所发展的基于FRET原理的双光子比率荧光探针具有显著特性：具有良好的发射光谱分辨率，可避免荧光光谱重叠而产生的严重荧光检测误差，有望为解决目前双光子比率荧光探针所面临的发射光谱重叠严重的问题提供理论指导和实验依据,具有重大的科学意义和应用价值。

本项目致力于具有优越光学性能的分子荧光探针开发，已进行了双光子小分子荧光探针的制备、光学性能和初步成像研究。从分子结构入手，发展了具有良好发射光谱分辨率的双光子荧光共振能量传递分子作为新型荧光探针平台，并基于该平台创建次氯酸、硫化氢和过氧化亚硝酰阴离子等的双光子比率荧光探针，实现了双光子比率成像检测组织与细胞内的各种生物活性物质。同时，课题组还在近红外荧光荧光染料平台构建及其应用于发展次氯酸等活性物质的分子荧光探针领域进行了系列研究。目前，该项目已取得一系列研究进展，发表了第一或通讯作者SCI论文9篇（IF>5的论文6篇），包括化学领域顶尖期刊J. Am. Chem. Soc、Angew. Chem.-Int. Ed和Chem. Sci论文各1篇，初步完成了项目设定的各项工作目标。