Hypochlorite (ClO−) plays an important role in many cellular processes, which is proved to be related to some serious diseases. Given the electron donating and electron withdrawing properties of benzo[h]chromene and rhodanine groups, respectively, we developed a Donor (D)-p-Acceptor (A) type probe 1 for the detection of ClO− based on benzo[h]chromene (D) and rhodanine (A), whose structure was determined by ESI-MS, NMR and X-ray diffraction. Probe 1 showed a fast (within 20 s), selective and sensitive response toward ClO− via a fluorescent turn-on output. Based-on the spectral change of probe 1 induced by ClO−, the mechanism associated with oxidation breakage of rhodanine moiety was proposed. Due to the excellent biocompatibility and low cytotoxicity, probe 1 can be used for imaging of ClO− in C6 cells, Arabidopsis thaliana, and zebrafish.