ABSTRACTA novel thermosensitive and water-soluble fluorescent polymer PNIPAM-curcumin (P2) containing curcumin unit was synthesized through one-pot method. The chemical structure and optical properties of compounds and polymer were characterized and tested. As the results turned out that P2 displayed excellent water solubility, thermosensitivity and exhibited strong fluorescence emission band in water. Moreover, P2 was further investigated by MTT assay and live cell imaging with HeLa cells. P2 did not have cytotoxicity in concentration of 200 µg · mL−1 within 48 h and the intracellular fluorescence intensity enhanced with the concentration of P2 varying from 0 to 500 µg · mL−1. According to the experimental results, as a fluorescence probe, P2 synthesized does not only possess nice water solubility but also has the advantage of no cytotoxicity, and it can easily enter the cells, which indicates that it is a characteristic of high sensitivity and responsiveness. Therefore, P2 will have immense potential to be applied in biomedical fields.