pH-Switchable Fluorescent Probe for Spatially-Confined Visualization of Intracellular Hydrogen Peroxide
pH-Switchable Fluorescent Probe for Spatially-Confined Visualization of Intracellular Hydrogen Peroxide
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pH 可切换荧光探针用于细胞内过氧化氢的空间限制可视化
DOI：
10.1021/acs.analchem.6b00654
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发表时间：
2016-06-07
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;Ren, Jing;Zhao, Yibing
细胞内H2O2在调节多种细胞功能中发挥着重要作用。能够对细胞内 H2O2 水平做出反应的荧光探针将为揭示 H2O2 在活生物体中的功能提供有价值的工具。然而，传统的 pH 不敏感探针和溶酶体靶向探针只能分别提供细胞内 H2O2 的空间非特异性可视化和溶酶体 H2O2 的特异性传感。在这项工作中，我们开发了一种 H2O2 响应和 pH 可切换的荧光探针 (HP-L1)，它可以对细胞内 H2O2 和溶酶体 pH 值依次做出响应。该荧光探针由 H2O2 响应的硼酸酯部分和 pH 可切换的螺苯并吡喃荧光团组成。当探针用于细胞内H2O2传感时，只有溶酶体的荧光发射可见，而其他区域的荧光无法明显检测到，这是由于螺苯并吡喃荧光团的pH可切换特性。因此，开发的荧光探针能够实现细胞内 H2O2 的空间限制（即溶酶体特异性）可视化。我们设想这种荧光探针（或提出的传感策略）将允许细胞内 H2O2 总体水平的可视化，而不会干扰来自其他来源（例如用于细胞染色和多重分析的染料）的可能荧光信号。
Intracellular H2O2 plays an important role in regulating a variety of cellular functions. Fluorescent probes that can make response to intracellular levels of H2O2 would provide valuable tools for revealing the functions of H2O2 in living organisms. However, traditional pH-insensitive probes and lysosome-targetable probes can only provide spatially nonspecific visualization of intracellular H2O2 and specific sensing of lysosomal H2O2, respectively. In this work, we developed a H2O2-responsive and pH-switchable fluorescent probe (HP-L1) which can make response sequentially to intracellular H2O2 and lysosomal pH. The fluorescent probe is comprised of a H2O2-responsive boronate moiety and a pH-switchable spirobenzopyran fluorophore. When the probe was applied for intracellular H2O2 sensing, only fluorescent emission from lysosomes is visible, and the fluorescence from other regions is not able to be obviously detected, which is due to the pH-switchable property of the spirobenzopyran fluorophore. Thus, the developed fluorescence probe enables the spatially confined (i.e., lysosome-specific) visualization of the intracellular H2O2. We envisioned that this kind of fluorescent probe (or the proposed sensing strategy) would allow the visualization of the overall levels of intracellular H2O2 without interferences of possible fluorescent signals from other sources (e.g., dyes for cellular staining and multiplex analysis).