Hypochlorite sensing and real-time imaging with XY-01: A red-emitting fluorescent turn-on probe for living cells and colorectal cancer organoids

Yichun Xu; Zhihua Chen; Jun Su; Yanting Ding; Jiajing Zhou; Jiawei Zhao; Zhiyuan He; Yi Gong; Zhai Cai; Lei Cui; Junsong Han

doi:10.17305/bb.2025.13312

Authors

Yichun Xu Department of Pathology, Shanghai Tongji Hospital, Tongji Hospital Affiliated to Tongji University, Shanghai, China; National Engineering Research Center for Biochip, Shanghai Biochip Limited Corporation, Shanghai, China. https://orcid.org/0000-0002-7339-1693
Zhihua Chen College of Science, Shanghai University, Shanghai, China
Jun Su Department of Pathology, Shanghai Tongji Hospital, Tongji Hospital Affiliated to Tongji University, Shanghai, China; National Engineering Research Center for Biochip, Shanghai Biochip Limited Corporation, Shanghai, China.
Yanting Ding Department of Pathology, Shanghai Tongji Hospital, Tongji Hospital Affiliated to Tongji University, Shanghai, China; National Engineering Research Center for Biochip, Shanghai Biochip Limited Corporation, Shanghai, China.
Jiajing Zhou National Engineering Research Center for Biochip, Shanghai Biochip Limited Corporation, Shanghai, China
Jiawei Zhao National Engineering Research Center for Biochip, Shanghai Biochip Limited Corporation, Shanghai, China
Zhiyuan He National Engineering Research Center for Biochip, Shanghai Biochip Limited Corporation, Shanghai, China
Yi Gong National Engineering Research Center for Biochip, Shanghai Biochip Limited Corporation, Shanghai, China
Zhai Cai Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou, China
Lei Cui College of Science, Shanghai University, Shanghai, China
Junsong Han National Engineering Research Center for Biochip, Shanghai Biochip Limited Corporation, Shanghai, China

DOI:

https://doi.org/10.17305/bb.2025.13312

Keywords:

Fluorescent probe, hypochlorite, organoid, cell imaging, near-infrared emission

Abstract

Hypochlorite (ClO⁻), a major reactive oxygen species generated in inflammation, is a potent biological oxidant involved in diverse physiological and pathological processes; therefore, sensitive detection of ClO⁻ is important for understanding disease pathophysiology and supporting early diagnosis and prevention. Here, we aimed to develop a physiologically compatible fluorescent tool for specific ClO⁻ sensing and imaging. We designed and synthesized a novel A–D–A type molecular fluorescent probe, XY-01, and characterized it by NMR, HRMS, UV–Vis and fluorescence spectroscopy. XY-01 operates through ClO⁻-triggered oxidation of a thioformyl group (C=S) to a carbonyl (C=O), which restores intramolecular charge transfer and produces a prominent fluorescence turn-on signal. In PBS (pH 7.4), XY-01 responded to ClO⁻ within 1 min with strong red emission at 666 nm and a large Stokes shift (~167 nm), showed high selectivity against common ions and reactive species, and achieved a detection limit of 3.39 µM within the biologically relevant range. Cytotoxicity assays indicated negligible toxicity, enabling real-time confocal imaging of ClO⁻ distribution in HCT-116 cells and colorectal cancer organoids. Collectively, XY-01 is a simple, sensitive, and low-toxicity probe that provides a promising platform for optical sensing and imaging of hypochlorite in living cells and organoids.

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