Time‐Resolved Luminescent Sensing and Imaging for Enzyme Catalytic Activity Based on Responsive Probes

Abstract Enzymes, as a kind of biomacromolecules, play an important role in many physiological processes and relate directly to various diseases. Developing an efficient detection method for enzyme activity is important to achieve early diagnosis of enzyme‐relevant diseases and high throughput screening of potential enzyme‐relevant drugs. Time‐resolved luminescence assay provide a high accuracy and signal‐to‐noise ratios detection methods for enzyme activity, which has been widely used in high throughput screening of enzyme‐relevant drugs and diagnosis of enzyme‐relevant diseases. Inspired by these advantages, various responsive probes based on metal complexes and metal‐free organic compounds have been developed for time‐resolved bioimaging and biosensing of enzyme activity owing to their long luminescence lifetimes, high quantum yields and photostability. In this review, we comprehensively reviewed metal complex‐ and metal‐free organic compound‐based responsive probes applied to detect enzyme activity through time‐resolved imaging, including their design strategies and sensing principles. Current challenges and future prospects in this rapidly growing field are also discussed.