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In recent years, fluorescent turn-on probes have become invaluable in diagnosing diseases and investigating pathological mechanisms due to their high sensitivity and minimal background interference. Among reactive oxygen species (ROS), hydrogen peroxide (H2O2) is particularly significant, as it plays a central role in regulating essential cellular processes like signaling, immune responses, and oxidative stress. In this research, we have produced an innovative fluorescent probe, HCyB, incorporating hemicyanine and arylboronate structures, specifically tailored for detecting H2O2 within biological systems. HCyB showed a robust linear fluorescence response to H2O2 concentrations ranging from 15 to 50 μM, with an impressive detection threshold of 76 nM, allowing for the identification of minute H2O2 levels. The probe exhibited exceptional selectivity for H2O2 over other biologically relevant molecules, reducing false signals and ensuring accurate detection. Additionally, HCyB demonstrated low cytotoxicity, making it ideal for live-cell imaging applications. Its ability to target mitochondria further increased its utility, given the critical role mitochondrial H2O2 plays in various cellular functions and disease mechanisms. The effectiveness of HCyB was confirmed across different cell lines, including mouse macrophages (RAW 264.7), human skin fibroblasts (WS1), breast cancer cells (MDA-MB-231), and human leukemia monocytic cells (THP1). HCyB successfully tracked both exogenous and endogenous H2O2 levels within these cells, highlighting its potential for real-time, in situ detection of H2O2 across various biological settings. This positions HCyB as a promising tool for biomedical research, particularly in studying oxidative stress-related diseases and H2O2-mediated cellular signaling pathways.

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A Targeted Fluorescent Probe for the Selective Detection and Imaging of Hydrogen Peroxide in Living Cells

  • 会议论文

  • 发表日期：

    2025年

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  • M. Aneeq U. Rehman,
  • Zhi-Jia Chen,
  • Lai Chian Hui

摘要

In recent years, fluorescent turn-on probes have become invaluable in diagnosing diseases and investigating pathological mechanisms due to their high sensitivity and minimal background interference. Among reactive oxygen species (ROS), hydrogen peroxide (H2O2) is particularly significant, as it plays a central role in regulating essential cellular processes like signaling, immune responses, and oxidative stress. In this research, we have produced an innovative fluorescent probe, HCyB, incorporating hemicyanine and arylboronate structures, specifically tailored for detecting H2O2 within biological systems. HCyB showed a robust linear fluorescence response to H2O2 concentrations ranging from 15 to 50 μM, with an impressive detection threshold of 76 nM, allowing for the identification of minute H2O2 levels. The probe exhibited exceptional selectivity for H2O2 over other biologically relevant molecules, reducing false signals and ensuring accurate detection. Additionally, HCyB demonstrated low cytotoxicity, making it ideal for live-cell imaging applications. Its ability to target mitochondria further increased its utility, given the critical role mitochondrial H2O2 plays in various cellular functions and disease mechanisms. The effectiveness of HCyB was confirmed across different cell lines, including mouse macrophages (RAW 264.7), human skin fibroblasts (WS1), breast cancer cells (MDA-MB-231), and human leukemia monocytic cells (THP1). HCyB successfully tracked both exogenous and endogenous H2O2 levels within these cells, highlighting its potential for real-time, in situ detection of H2O2 across various biological settings. This positions HCyB as a promising tool for biomedical research, particularly in studying oxidative stress-related diseases and H2O2-mediated cellular signaling pathways.