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The effect of Epimedium extract (EE) on erythrocytes was investigated by atomic force microscopy (AFM). The images of the surface structures showed clear concave and progressive increase of surface roughness of erythrocyte after incubation with EE at concentration of 0.2 or 0.05 g/L, far below its critical hemolytic levels. The AFM results also indicated that the granules of the fine surface structure increased, which caused by aggregation of membrane protein. Further study showed that the change in surface topography of erythrocyte membrane might be connected with the increase of intracellular free Ca2+ induced by EE.