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An “off-on” fluorescent nanoprobe for near-infrared multiphoton imaging of singlet oxygen has been developed. The nanoprobe comprises a naphthoxazole fluorescent unit and a singlet-oxygen-sensitive furan derivative attached to the surface of mesoporous silica nanoparticles. In solution, the fluorescence of the nanoprobe increases upon reaction with singlet oxygen both under one- and multiphoton excitation, with fluorescence enhancements up to 180-fold. The nanoprobe can be readily internalized by macrophage cells and is capable of imaging intracellular singlet oxygen under multiphoton excitation.