新型双光子比例计量型锌离子荧光探针的构筑

项目名称： 新型双光子比例计量型锌离子荧光探针的构筑

项目编号： No.20871066

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 何卫江

作者单位： 南京大学

项目金额： 33万元

中文摘要： 为克服目前增强型锌离子荧光探针易受环境和探针浓度影响的不足，本项目主要通过对ICT荧光团的推或拉电子基进行化学修饰为锌离子螯合团，结合推拉电子能力的调节来构建比例计量型锌离子荧光探针。首先利用锌离子配位诱导荧光团共面化结合ICT效应调节的策略构建了基于2-吡啶基苯并噁唑荧光团的比例计量型锌探针DBITA。该探针显示锌特异诱导的荧光发射红移(由534到609 nm)，细胞造影证实了其锌离子比例造影能力。同时通过增加2,1,3-苯并噁二唑的推电子基中的锌配位数、降低拉电子基的拉电子能力，分别获得了可见光激发比例计量型锌探针：SBD-TPE和SHNT。利用前者不仅实现了活细胞锌的比例造影，证实了其高尔基体分布特点，实现了高尔基体锌浓度的定量，而且通过活体斑马鱼头部锌离子的比例成像，实现了活体锌离子的定量检测。最后通过BODIPY荧光团的单侧TPEA衍生化构建了近红外比例计量型锌离子荧管探针BDP-TPEA。该探针显示了锌离子特异的激发蓝移的比例响应特点（570到512 nm），其近红外发射的优势帮助实现了活体裸鼠的锌离子比例成像，为锌离子定量成像进入临床辅助诊断奠定了基础。

中文关键词： 锌离子;荧光探针;造影;细胞;斑马鱼

英文摘要： This project focused on the development of ratiometric Zn2+ sensor to overcome the disadavatages od MCHEF-tyepe sensors, such as the microenvironment- and concentration dependent sensing behavior, by modifying the donor(D) or acceptor (A) of ICT fluorophores as zinc ionophore and A/D tuning to acquire the Zn2+ ratiometric sensing behavior.DBITA was developed at first by a strategy of Zn2+-chelation induced coplanation of 2-(2-pyridyl)benzoxazole fluorophore.DBITA exhibits the specific Zn2+-induced emission bathochromic shift from 534 to 609 nm, and cell imaging on HeLa cells confirmed its ratiometric imaging ability for intracellaur Zn2+.Then two new visible light excitable ratiometric Zn2+ sensors SBD-TPEA and SNHT were constructed from 2,1,3-benzoxadiazole via enhancing the synergic Zn2+ coordination number in D and decreasing the electron-withdrawing ability of A. Cellular imaging with SBD-TPEA verfied not only its ratiometric Zn2+ imaging ability but also the Golgi-preferential affinity, and Golgi Zn2+ has been quantified in the imaging. In addition,in vivo ratiometric Zn2+ imaging was firstly realized with SBD-TPEA on zebrafish larva and Zn2+ quantification on larva head has been finished.Finally,a near infrared (NIR) ratiometric Zn2+ sensor BDP-TPEA was prepared based on the mono-TPEA modification of BODIPY fluorophore. It displayed the Zn2+-induce excition hypsochromic shift from 570 to 512 nm.Its NIR emssion-favored in vivo Zn2+ ratimetric imaging in mice should be helpful to develope clinical technique to assist the diagnosis of Zn2+ disorder-related diseases.

英文关键词： zinc cation; fluorescent sensor;imaging;cell; zebrafish