Simultaneous and non-destructive quantitative detection of intracellular metal ions holds great promise for improving the accuracy of diagnosis and biological research. Herein, novel multicolor DNAzymes-embedded framework nucleic acids (FNAzymes) were presented, which can easily enter cells and achieve simultaneous and quantitative detection of intracellular physiologically related Cu2+ and Zn2+. Two types of DNAzymes, specific to Cu2+ and Zn2+, were encoded in the framework nucleic acids (FNAs) via self-assembly. With the formation of a well-ordered FNAzyme nanostructure, the fluorophore and the quencher were close to each other; therefore, the fluorescence was quenched. In the presence of Cu2+ and Zn2+, the integrated FNAzymes would be specifically cleaved, resulting in the release of fluorophores in cells. Consequently, the fluorescence in living cells could be observed by a confocal microscope and semi-quantitatively analyzed by flow cytometry with low-nanomolar sensitivity for both metal ions. The FNAzymes have high uniformity and structural accuracy, which are beneficial for intracellular detection with excellent reproducibility. This proposed method offers new opportunities for non-destructive, semi-quantitative, multi-target detection in living cells.
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