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Benzopyran nitrile dyes cannot be used as qualified photosensitizers due to the low quantum yield of triplet state. The benzopyran derivatives containing selenium instead of oxygen atom based on the heavy atom effect are expected to become potential agents for photodynamic therapy. In this paper, a series of selenium-containing photosensitizers (PS X) were prepared according to this strategy. PS X can effectively produce both singlet oxygen and superoxide anions upon laser irradiation. PS X exhibited the emission wavelength at 500–800 nm and near-infrared (NIR) fluorescence imaging in HeLa cells. Excellent biocompatibility and phototoxicity further indicated that PS X could be used as efficient photosensitizers for NIR fluorescence imaging and photodynamic therapy. Four novel selenium-containing photosensitizers (PS X) have been synthesized through a strategy of introducing Se into the dye core. PS X can serve as effective photosensitizers for NIR fluorescence imaging and photodynamic therapy. [Display omitted] • Four selenium-containing photosensitizers (PS X) were prepared by introducing Se into the dye core. • PS X can effectively produce both singlet oxygen and superoxide anions upon laser irradiation. • PSx exhibited near-infrared fluorescence imaging and efficient photodynamic therapy in HeLs cells. [ABSTRACT FROM AUTHOR]