Mechanistic insights into electrochemiluminescence: fueling advances in bioanalysis and imaging.

The rational improvement of electrochemiluminescence (ECL) features such as increased signal intensity and both spatial and temporal extension requires deciphering the competitive mechanistic pathways occurring in the vicinity of the electrode surface. This understanding is critical for developing high-performance ECL analytical systems. This perspective systematically reviews the advanced characterization and theoretical simulation techniques for uncovering ECL reaction mechanisms, including ECL microscopy, ECL-scanning electrochemical microscopy, ECL self-interference spectroscopy, electrochemistry-mass spectrometry, and ECL-voltammetric analysis and numerical simulation. Emerging ECL enhancement mechanisms and regulatory pathways are comprehensively elaborated, covering peroxydisulfate-oxalate and redox-mediated mechanisms, time-scale regulation strategies of radical stabilization, time matching, and pre-oxidation/reduction, as well as exogenous stimulation modulation via heat, ultrasound, and light. Moreover, this article highlights recent progress in the application of enhanced ECL platforms in bioanalysis and bioimaging. This review offers a systematic and instructive reference for the rational design and further development of high-efficiency ECL systems for advanced bioanalysis and microscopy applications.