A Doubly-Quenched Fluorescent Probe for Low-Background Detection of Mitochondrial H2O2
A Doubly-Quenched Fluorescent Probe for Low-Background Detection of Mitochondrial H2O2
复制标题
用于线粒体 H2O2 低背景检测的双猝灭荧光探针
DOI：
10.1021/acs.analchem.9b01294
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发表时间：
2019-05-21
影响因子：
7.4
通讯作者：
Zhao, Yibing
中科院分区：
文献类型：
--
作者：
Liu, Jun;Liang, Jingjing;Zhao, Yibing
过氧化氢（H2O2）是氧代谢的重要产物，在调节多种细胞功能中起着至关重要的作用。荧光探针为我们理解内源性H2O2的生物学作用做出了巨大贡献。然而，具有芳基硼酸酯的H2O2的荧光探针可遭受中等开启荧光响应。策略，可以减少这些硼酸盐掩蔽的探针的背景荧光将显着提高内源性过氧化氢检测的灵敏度。在这项工作中，我们提出了一个通用的和可靠的双淬灭的概念，低背景荧光的荧光探针的设计。本文报道了一种新的荧光探针，用于检测肝癌细胞线粒体内源性H_2O_2。该探针同时利用硼酸盐驱动的内酰胺形成和可消除的淬灭部分（即，双猝灭效应）以降低背景荧光，这最终导致实现>50倍的荧光开启。可以获得在1和60 μ M之间的响应的线性浓度范围和0.025 μ M的检测限。这项研究不仅提出了一个高灵敏度的荧光探针检测过氧化氢，但也提供了一个新的概念，荧光探针的设计与以前无法实现的其他类型的活性氧和许多其他生物相关的分析物的荧光开关响应比。
Hydrogen peroxide (H2O2) is an important product of oxygen metabolism and plays a crucial role in regulating a variety of cellular functions. Fluorescent probes have made a great contribution to our understanding of the biological role of endogenous H2O2. However, fluorescent probes for H2O2 featuring aryl boronates can suffer from moderate turn-on fluorescence responses. Strategies that can reduce the background fluorescence of these boronate-masked probes would significantly improve the sensitivity of endogenous H2O2 detection. In this work, we propose a general and reliable double-quenching concept for the design of fluorescent probes with low background fluorescence. A new fluorescent probe was developed for the detection of endogenous H2O2 in mitochondria of live cancer cells. This probe exploits a boronate-driven lactam formation and an eliminable quenching moiety simultaneously (i.e., the double-quenching effect) to reduce the background fluorescence, which ultimately results in the achievement of a >50-fold fluorescence turn-on. A linear concentration range of response between 1 and 60 mu M and a detection limit of 0.025 mu M can be obtained. This study not only presents a highly sensitive fluorescent probe for the detection of H2O2 but also provides a new concept for the design of fluorescent probes with a previously unachievable fluorescence off-on response ratio for other types of ROS and many other biologically relevant analytes.