A Schiff-based fluorescent molecular probe (HSZn) was synthesized by condensation of quinoline with phenylhydrazine and characterized by FTIR, NMR and HRMS. The UV-Vis spectroscopy and fluorescence spectroscopy studies demonstrated that the probe showed a significant fluorescence enhancement effect toward Zn2+ in aqueous medium while suffering from a slight interference at the presence of other metal ions. A good linear relationship between the Zn2+ concentrations (1.0×10-5~1.0×10-4 mol/L) and the fluorescence intensity was obtained for determining the content of Zn2+ with a detection limit of 4.0×10-8 mol/L. A 1:2 binding ratio of Zn2+/HSZn for the complex was obtained by Job’s plot and LC-MS, HSZn also exhibited a sensitive fluorescence response toward Zn2+ in the wide pH range (pH=5~13). Cell imaging studies indicated that the probe was capable of detecting endogenous Zn2+ in live Hela cells.