Fluorescent Probing of Hydrogen Peroxide in Biological Systems

Hydrogen peroxide (H₂O₂) serves as both a signalling molecule in physiological processes and an indicator of oxidative stress in disease states. Its transient nature and subcellular compartmentalisation demand highly sensitive and selective imaging tools. Fluorescent probes—ranging from small synthetic molecules to genetically encoded reporters—have emerged as indispensable for visualising H₂O₂ dynamics in living cells and tissues. Advances in probe chemistry now enable ratiometric measurements, dual‐colour or multi‐mode detection, organelle targeting and compatibility with deep‐tissue microscopy. Such innovations allow real‐time tracking of H₂O₂ generation and clearance during signalling events, apoptosis, inflammation and ageing. The integration of novel photophysical strategies—including aggregation‐induced emission and upconversion nanotechnology—has further enhanced detection limits and spatial resolution. Together, these approaches provide a powerful toolkit to dissect the roles of H₂O₂ in health and disease and to accelerate the development of diagnostic and therapeutic applications.

Research from Nature Portfolio

Recent studies have introduced a ratiometric fluorescent probe exhibiting aggregation‐induced emission properties, which offers well‐resolved dual emission peaks upon reaction with endogenous H₂O₂. This probe displays high sensitivity, excellent selectivity against other reactive species, low cytotoxicity and efficient cell‐membrane permeability. It has enabled the first comparative imaging of H₂O₂ levels in macrophages and cancer cells, revealing substantially higher peroxide concentrations in malignant cells. The work demonstrates the potential of aggregation‐induced emission platforms for precise, real‐time quantitative mapping of oxidative signals in diverse biological contexts.

Fluorescent Probing of Hydrogen Peroxide in Biological Systems publication trend

The graph below shows the total number of articles in fluorescent probing of hydrogen peroxide in biological systems across all publications each year (not limited to Nature Index journals).

Technical terms

Fluorescent probe: Molecule that emits light upon excitation, allowing in situ detection of specific analytes.

Ratiometric imaging: Method measuring fluorescence at two wavelengths to correct for probe concentration and environmental variability.

Aggregation‐induced emission (AIE): Phenomenon where fluorophores become highly emissive upon aggregation, enhancing brightness in biological settings.

Boronate‐based probe: Compound bearing a boronate group that reacts selectively with H₂O₂ to produce a fluorescent product.

Upconversion nanoparticles (UCNPs): Materials converting near‐infrared excitation into visible emission, enabling deep‐tissue imaging with low autofluorescence.

References

1. A Self‐Accelerating Naphthalimide‐Based Probe Coupled with Upconversion Nanoparticles for Ultra‐Accurate Tri‐Mode Visualization of Hydrogen Peroxide. Advanced Science (2024).
2. Ratiometric fluorescent probe with AIE property for monitoring endogenous hydrogen peroxide in macrophages and cancer cells. Scientific Reports (2017).
3. A Novel Fluorescent Probe for Hydrogen Peroxide and Its Application in Bio-Imaging. Molecules (2021).
4. Boronate-Based Probes for Biological Oxidants: A Novel Class of Molecular Tools for Redox Biology. Frontiers in Chemistry (2020).