The rapid evolution of antibiotic resistance increasingly challenges the successful treatment of S. aureus infections. Here, we present an unconventional treatment approach by disassembly its membrane microdomains via pulsed laser photolysis of staphyloxanthin. After staphyloxanthin photolysis, membrane permeabilization, fluidification, and membrane protein detachment, were found the underlying mechanisms to malfunction its defense to several major classes of conventional antibiotics. Through resistance selection study, we found pulsed laser treatment completely depleted staphyloxanthin virulence. More importantly, laser treatment further inhibited development of resistance for several major classes of conventional antibiotics including fluoroquinolones, tetracyclines, aminoglycosides, and oxazolidinones. Collectively, this work highlights a novel platform to revive conventional antibiotics to treat S. aureus infections.