Recent progress in chemiluminescence techniques for reactive oxygen species detection

Reactive oxygen species (ROS) play crucial roles in a wide range of physiological and pathological processes. Consequently, their efficient and selective detection is of significant importance for disease diagnosis, therapeutic monitoring, and environmental analysis. Chemical reaction–driven luminescence techniques, including chemiluminescence (CL), bioluminescence (BL), and electrochemiluminescence (ECL), have emerged as powerful detection modalities, offering high sensitivity, intrinsic background suppression, and operation without external light excitation, thus minimizing photodamage and autofluorescence. Over the past five years, these platforms have undergone rapid innovation in molecular design, materials integration, and system engineering, dramatically expanding their capabilities for ROS sensing in complex biological environments. In CL systems, advanced architectures incorporating nanomaterials, polymeric matrices, and hybrid composites have yielded substantial gains in analytical performance. These enhancements have increased photon yield, extended operational stability, and enabled multifunctional sensing–imaging platforms. The development of near-infrared (NIR) CL probes has been particularly transformative, affording deeper tissue penetration and improved in vivo imaging resolution. BL approaches have capitalized on resonance energy transfer cascades and activatable probe chemistries to achieve highly selective, low-background imaging of specific ROS species, facilitating longitudinal monitoring of oxidative events in living systems. ECL strategies have progressed through the synthesis of novel luminophore-doped nanostructures and the engineering of functionalized electrode interfaces, enhancing signal responsiveness, operational stability, and biocompatibility. Notably, flexible and bio-integrated ECL devices are emerging, enabling conformal interfacing with biological tissues for localized ROS sensing. Despite these advances, fundamental challenges persist. CL detection remains vulnerable to environmental fluctuations in pH and temperature, constraining stability in vivo. BL probes often suffer from poor substrate stability, limited penetration depth, and the risk of perturbing normal cellular metabolism. ECL typically requires high voltages or non-physiological electrolytes, which limit direct in vivo applications; additionally, its selectivity for distinct ROS species is often insufficient, with signals susceptible to interference from a wide range of redox-active biomolecules. Overcoming these limitations demands innovation in molecular probe chemistry, adaptive materials, and system-level integration to enable simultaneous multi-ROS detection, robust biological stability, and theranostic functionality. Looking ahead, the convergence of chemical luminescence technologies with cutting-edge innovations is poised to redefine ROS sensing. Future research is expected to focus on smart-responsive mechanisms capable of adapting to fluctuating biological microenvironments, seamless integration of multi-modal detection platforms, and AI-assisted data analysis to enhance signal interpretation and predictive modeling. These advances aim to propel ROS detection toward high-throughput screening, real-time visualization of redox dynamics, and precision medicine applications. The design of next-generation luminescent substrates with superior photophysical stability, optimized quantum yield, and tunable emission wavelengths, combined with synergistic integration with complementary imaging or biosensing modalities, will further elevate spatial resolution, molecular specificity, and background discrimination. Such progress will not only deepen mechanistic understanding of ROS-mediated signaling and pathology but also accelerate translation into clinically actionable diagnostics and targeted interventions.