[Objective] Essential oil (EO) composition of Coreopsis tinctoria Nutt., together with The their anti-Cryptococcus activity and influence on the cell membrane of Cryptococcus neoformans present study were evaluated in this study. [Methods] EO from Coreopsis tinctoria Nutt. was extracted team distillation and the main chemical components of it was identified by GC-MS. The micro-dilution method was used in acquiring the minimal inhibitory concentration (MIC). The cell weight of C. neoformans and germ tube germination of C. neoformans were also determined to evaluate the anti-C. neoformans activity in vitro. The effect on membrane permeabilization was measured by monitoring the influx of Propidium iodide (PI) using flow cytometry and further verified through the inhibition of ergosterol synthesis in the cell membrane. [Results] The MIC value of EO against C. neoformans was 0.781 μL/mL. Cell weight and germ tube germination of C. neoformans were inhibited in a dose-dependent manner. As shown in the results ergosterol production was reduced and propidium iodide penetrated C. neoformans through a lesion in its cell membrane upon treatment with EO. [Conclusion] Inhibitory effect of EO against C. neoformans relying on the cell membrane lesion.