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Engineering a Reversible Fluorescent Probe for Real-Time Live-Cell Imaging and Quantification of Mitochondrial ATP

机译:工程一种可逆荧光探针,用于实时活细胞成像和线粒体ATP的定量

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摘要

Real-time imaging and quantification of adenosine triphosphate (ATP) fluctuation in cells are significant for understanding the relationship between energy metabolism and cell functions. However, few synthetic fluorescent probes have been reported to tackle this challenge due to lack of accurate fluorescence readout and suitable response concentration. Herein we designed and synthesized a ratiometric fluorescent probe (Rh6G-ACFPN) for quantitatively detecting the fluctuation of mitochondrial ATP in living cells. Rh6G-ACFPN selectively and reversibly responds to ATP with an ideal dissociation constant (K-d) of 4.65 mM (3-10 mM: the range of mitochondrial ATP concentrations). Live-cell imaging allows us to directly monitor the dynamic changes of mitochondrial ATP in high temporal resolution. Moreover, for the first time, mitochondrial ATP in normal and cancer cells lines was successfully quantified and discriminated. These results demonstrate the versatility of Rh6G-ACFPN as a useful imaging tool to elucidate the function of mitochondrial ATP in living cells.
机译:细胞三磷酸三磷酸腺苷(ATP)波动的实时成像和定量对于了解能量代谢和细胞功能之间的关系很重要。然而,由于缺乏精确的荧光读数和合适的反应浓度,据报道,据据报道,少数合成的荧光探针在缺乏准确的荧光读数和合适的响应浓度,以解决这一挑战。在此,我们设计并合成了用于定量检测活细胞中线粒体ATP的波动的比例荧光探针(RH6G-ACFPN)。 RH6G-ACFPN选择性地和可逆地响应ATP,理想的解离常数(K-D)为4.65 mm(3-10mm:线粒体ATP浓度的范围)。 Live-Cell Imaging允许我们直接监测高时间分辨率的线粒体ATP的动态变化。此外,首次,正常和癌细胞系中的线粒体ATP被成功地量化和鉴别。这些结果证明了RH6G-ACFPN作为一种有用的成像工具,以阐明活细胞中线粒体ATP的功能。

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  • 来源
    《Analytical chemistry》 |2020年第6期|共8页
  • 作者单位

    Hunan Univ Coll Chem &

    Chem Engn State Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn State Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Max Planck Inst Med Res Dept Chem Biol D-69120 Heidelberg Germany;

    Hunan Univ Coll Chem &

    Chem Engn State Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn State Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn State Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn State Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分析化学;
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