A mitochondria-targeted phosphorescent probe for sequentially detecting Cu2+ and cysteine and its imaging in living cells and in vivo

This study reported a unique mitochondria-targetable phosphorescent 'on-off-on' probe, Ir-DPA, based on an iridium(III) complex derivative for the sequential and specific detection of Cu2+ and Cys in vitro and in vivo. Probe Ir-DPA was constructed by a lipophilic cation unit as the chromophore as well as the mitochondria biomarker and a dipicolylamine (DPA) group as the recognition unit for Cu2+, followed by removal of Cu2+ specifically in the presence of Cys. The probe itself was strongly emissive at 570 nm. Upon addition of Cu2+, marked quenching occurred as result of the binding of Cu2+ with DPA moiety of this probe, accompanied by a distinct naked-eye luminescence color change from orange yellow to colorless. Additionally, further addition of Cys, the displacement of Cu2+ from in situ generated Ir-DPA-Cu ensemble because of the large nucleophilicity of cysteine toward Cu2+ led to the release of probe, giving rise to the luminescence recovery, as well as a relevant chromogenic change from colorless to orange yellow. Ir-DPA showed a large Stokes shift of 200 nm and high selectivity and sensitivity for the rapid detection of Cu2+ and Cys under physiological conditions, with low detection limits of 0.04 μM and 1.21 μM for Cu2+ and Cys, respectively. More importantly, the probe successfully targeted mitochondria, sensitively detected Cu2+ and exogenously or endogenously generated Cys both in mitochondria of living cells and zebrafish. Finally, Ir-DPA was capable of quantifying Cu2+ level in water samples and the in situ generated ensemble identifying Cys in human serum. These results indicated the great potential of Ir-DPA for monitoring Cu2+ and Cys in environmental, biological systems, and mitochondrial processes. A mitochondria-targetable phosphorescent 'on-off-on' probe for sequentially and selectively detecting Cu2+ and cysteine has been achieved based on Cu2+-displacement reaction. [Display omitted] •A mitochondria-targetable phosphorescent 'on-off-on' probe for the specific detection of Cu2+ and cysteine was developed.•The probe exhibited a sequential orange yellow-colorless-orange yellow phosphorescent response to Cu2+ and cysteine.•The probe was successfully used for imaging Cu2+ and cysteine both in mitochondria of living cells and animals.