Highly Selective Two-Photon Fluorescent Probe for Ratiometric Sensing and Imaging Cysteine in Mitochondria
Highly Selective Two-Photon Fluorescent Probe for Ratiometric Sensing and Imaging Cysteine in Mitochondria
复制标题
用于线粒体中半胱氨酸比例传感和成像的高选择性双光子荧光探针
DOI：
10.1021/acs.analchem.5b04329
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发表时间：
2016-02-02
影响因子：
7.4
通讯作者：
Wong, Man Shing
中科院分区：
文献类型：
--
作者：
Niu, Weifen;Guo, Lei;Wong, Man Shing
以巯基为荧光团，丙烯酸酯基团为巯基反应位点，设计了一种新型的线粒体半胱氨酸（Cys）双光子荧光探针。生物相容性和光稳定性丙烯酸酯功能化的mercury探针不仅显示出一个cathetargeting属性，但也高度选择性检测和监测其他生物硫醇，如同型半胱氨酸（Hcy）和谷胱甘肽（GSH）和硫化氢（H2S）在活细胞中的Cys。此外，该探针显示比率荧光发射特性（F-518/F-452），其在0.5-40 μ M范围内与Cys浓度成线性比例。更重要的是，探针及其释放的荧光团mercury，表现出强的双光子激发荧光（TPEF）与双光子作用截面（Phi sigma（max））分别为65.2 GM在740 nm和72.6 GM在760 nm的水介质中，这是非常理想的高对比度和亮度比双光子荧光成像的活样品。该探针已成功地应用于比率图像和检测线粒体半胱氨酸在活细胞和完整组织的深度为150 μ m的双光子荧光显微镜。因此，这种比率双光子荧光探针实际上是有用的半胱氨酸在活的生物系统中的调查。
A novel ratiometric mitochondrial cysteine (Cys)-selective twophoton fluorescence probe has been developed on the basis of a merocyanine as the fluorophore and an acrylate moiety as the biothiol reaction site. The biocompatible and photostable acrylate-functionalized merocyanine probe shows not only a mitochondriatargeting property but also highly selective detection and monitoring of Cys over other biothiols such as homocysteine (Hcy) and glutathione (GSH) and hydrogen sulfide (H2S) in live cells. In addition, this probe exhibits ratiometric fluorescence emission characteristics (F-518/F-452), which are linearly proportional to Cys concentrations in the range of 0.5-40 mu M. More importantly, the probe and its released fluorophore, merocyanine, exhibit strong two-photon excited fluorescence (TPEF) with two-photon action cross-section (Phi sigma(max)) of 65.2 GM at 740 nm and 72.6 GM at 760 nm in aqueous medium, respectively, which is highly desirable for high contrast and brightness ratiometric two-photon fluorescence imaging of the living samples. The probe has been successfully applied to ratiometrically image and detect mitochondrial Cys in live cells and intact tissues down to a depth of 150 mu m by two-photon fluorescence microscopy. Thus, this ratiometric two-photon fluorescent probe is practically useful for an investigation of Cys in living biological systems.