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A bifunctional nanoparticles-based carrier for simultaneous in vivo imaging and photodynamic therapy by encapsulating methylene blue (MB) alone in the phosphonate-terminated silica matrix has been developed. The phosphonate-terminated silica nanoparticles, entrapping water-soluble photosensitizer MB (MB-encapsulated PSiNPs), are synthesized by the controlled synchronous hydrolysis of tetraethoxysilane and trihydroxyl silyl propyl methyl phosphonate in the water-in-oil microemulsion. The resulting MB-encapsulated PSiNPs effectively prevent the leakage of entrapped MB from the particles and provide protection for against reduction by diaphorase. Enough dose of irradiation to the MB-encapsulated PSiNPs under the light of 635 nm results in efficient generation of singlet oxygen and induces photodynamic damage to Hela cells. Furthermore, the non-invasive visualization of MB-encapsulated PSiNPs in …