Bioluminescence Recovery after Photobleaching (BRAP) Enables Real-Time Imaging of Self-Repair Dynamics and Discovery...

Self-repair is a fundamental biological process that not only maintains homeostasis but also drives drug resistance in pathogens and tumors. Conventional methods to quantify repair rely on cell proliferation, making them incompatible with nondividing cells─such as neurons and dormant microbes, and often require impractically long assay times. We present bioluminescence recovery after photobleaching (BRAP), a chemical biology platform that enables real-time, on-demand imaging of self-repair dynamics in living organisms. Using bioluminescent bacteria as a model, BRAP quantifies repair activity within 1-2 h and allows high-throughput screening of repair modulators. Among 20 amino acids, l-serine most strongly promoted repair, functioning stereospecifically over d-serine. Antibiotic screening further revealed that self-repair inhibition did not directly correlate with bactericidal efficacy. Tetracycline and gold(III) complexes were identified as effective repair suppressors operating primarily through reactive oxygen species (ROS) induction. BRAP represents a versatile and efficient chemical tool for directly probing repair mechanisms and accelerating the discovery of modulators against antibiotic resistance or for cellular recovery applications.