Dual-channel fluorescent probes enable synchronous tracking of peroxynitrite and cysteine in mitochondrial redox dynamics.

Authors

Chunyan Liu, Danyi He, Yanfei Lin, Tianqing Ye, Zhaosheng Qian, Haiying Liu, Longhua Guo, Jianbo Wang

Published in

Talanta. Volume 311. Pages 130253. Jul 01, 2026. Epub Jul 01, 2026.

Abstract

Peroxynitrite (ONOO^-) and cysteine (Cys) are pivotal components of reactive nitrogen and sulfur species that together shape a complex redox regulatory network in living systems. High-resolution, real-time imaging of these species is essential for unraveling their roles in mitochondrial redox dynamics. However, the development of fluorescent probes capable of simultaneously detecting ONOO^- and Cys with high fidelity remains limited. In this work, we introduce a single-excitation, dual-response fluorescent probe system (probes A and B) that enables synchronous visualization of ONOO^- and Cys. The probe design integrates two orthogonal sensing chemistries: ONOO^- is selectively recognized via the oxidation of a naphthylimide-borate unit, while Cys is detected through nucleophilic substitution with a benzothiol-modified tetrahydro-acridine conjugate salt. Incorporation of a long-chain linker effectively suppresses undesired FRET, and the large Stokes shift of the Cys-responsive unit ensures clean spectral separation. As a result, probe B exhibits strong ONOO^- emission at 550 nm and distinct Cys emission at 652 nm under a single 460 nm excitation source, with both fluorescence channels operating independently and without spectral crosstalk. Probe B demonstrates high sensitivity, excellent anti-interference capability, and precise mitochondrial localization. Dual-channel imaging in living cells confirms its ability to simultaneously monitor endogenous ONOO^- and Cys. Importantly, the probe enables real-time visualization of dynamic redox fluctuations during inflammatory responses and tumor ferroptosis. Overall, this work presents a robust dual-channel fluorescent imaging platform for the simultaneous detection of ONOO^- and Cys, offering a powerful tool for dissecting mitochondrial redox dynamics in health and disease.

PMID:
42402224
Bibliographic data and abstract were imported from PubMed on 06 Jul 2026.

Read full publication at:
Please sign in to see all details.

Stats

1-terrible, 9-excellent. How would you rate this publication? Sign in in to submit your rating.

Post a comment

You need to be signed in to post comments. You can sign in here.

Comments

There are no comments yet.