Dibenzylfluorescein is widely used as a fluorescent probe in biochemical and cellular research. Its primary application lies in detecting and measuring enzymatic activity, particularly in assays involving esterases and lipases. The compound is non-fluorescent in its intact form but becomes highly fluorescent upon enzymatic cleavage, making it a valuable tool for tracking enzyme kinetics and activity in real-time. Additionally, it is employed in imaging studies to visualize cellular processes and monitor intracellular enzyme distribution. Its sensitivity and specificity make it a preferred choice in high-throughput screening and diagnostic applications.