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A method was developed to determine the changes of the Arabidopsis thaliana tissues membrane potentials rapidly. Arabidopsis thaliana stem epidermal cells were labeled by different concentrations of the hydrophobic fluorescence dye DiBAC4(3), and the changes of the membrane potentials were measured dynamically by Confocal laser scanning microscope (CLSM). The effects of Jasmonic acid on Arabidopsis thaliana stem epidermal cells membrane potentials were investigated by the established method. The results showed that the best effect is the DiBAC4 (3) final concentration of 5 μmol·L-1.