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基于功能 DNA 自组装的信号放大实现活细胞中 Zn 和 Cu 的高灵敏荧光成像。

Highly Sensitive Fluorescence Imaging of Zn and Cu in Living Cells with Signal Amplification Based on Functional DNA Self-Assembly.

机构信息

College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging, Institute of Molecular and Nano Science , Shandong Normal University , Jinan , 250014 Shandong , People's Republic of China.

出版信息

Anal Chem. 2018 Aug 7;90(15):8785-8792. doi: 10.1021/acs.analchem.7b05268. Epub 2018 Jul 18.

Abstract

Intracellular trace Zn and Cu play important roles in the regulation of cell function. Considering the limitations of existing metal ion detection methods regarding sensitivity and applicability to living cells, an amplification strategy based on functional DNA self-assembly under DNAzyme catalysis to improve the sensitivity of intracellular Zn and Cu imaging is reported. In this process, metal ions as cofactor can activate the catalysis of DNAzyme to shear substrate chains, and each broken substrate chain can initiate consecutive hybridizations of hairpin probes (Hx) labeled with fluorophore, which can reflect the information on a single metal ion with multiple fluorophores. The detection limit can reach nearly 80 pM and high-sensitivity fluorescence imaging of intracellular Zn and Cu can be achieved. The results are important for research on cell function regulation associated with trace Zn and Cu. This approach is also a new way to improve the sensitivity of other trace metal ion imaging.

摘要

细胞内痕量的 Zn 和 Cu 在调节细胞功能方面发挥着重要作用。考虑到现有金属离子检测方法在灵敏度和适用于活细胞方面的局限性,本研究报道了一种基于在 DNA 酶催化下的功能 DNA 自组装的放大策略,以提高细胞内 Zn 和 Cu 成像的灵敏度。在这个过程中,金属离子作为辅助因子可以激活 DNA 酶的催化作用,剪切底物链,每个断裂的底物链都可以引发带有荧光团标记的发夹探针(Hx)的连续杂交,从而可以通过多个荧光团反映单个金属离子的信息。检测限可达到近 80 pM,并可实现对细胞内 Zn 和 Cu 的高灵敏度荧光成像。这些结果对于研究与痕量 Zn 和 Cu 相关的细胞功能调节具有重要意义。该方法也是提高其他痕量金属离子成像灵敏度的新途径。

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