A methylene blue (MB)-based beta-galactosidase (β-gal) activatable molecule, Gal-MB, was developed for senescence imaging and light-triggered senolysis. When in contact with LacZ β-gal or senescence-associated β-gal (SA-β-gal), the photoinsensitive Gal-MB becomes fluorescent. Gal-MB also offered selective phototoxicity toward LacZ β-gal expressing cells and drug-induced senescent cells, which express SA-β-gal, after light illumination at 665 nm.