Persisters, which are characterized by their multidrug tolerance, are phenotypic variants of regular microorganisms. Evidence indicates that persisters may play an important role in the recalcitrance of chronic infections, but the mechanism of persister formation is not yet clear. Although it is known that Candida albicans persisters can form only in biofilm environment, the identity of the biofilm element(s) that trigger persister formation are unknown. In this study, we tried to know the possible relationship between C. albicans persister formation and surface adhesion. We tested C. albicans persister formation dynamically at different time points during the process of adhesion and biofilm formation on the surface of biomaterials (polystyrene). Immediately following adhesion of C. albicans to the surface, persister cells emerged, and the proportion of persisters reached a peak of ~1% in approximately 2 h biofilm. As the biofilm matured, the proportion of persisters decreased and was only 0.01% by 24 h, while the number of persisters remained stable with no significant change. Persisters were not detected in the absence of an attachment surface pre-coated with C. albicans CAI-4; they were also absent when adhesion was destroyed by scraping the biofilm away from attachment surface, before treatment with antifungal drug. These results indicate that surface adhesion is closely related with the emergence and maintenance of Candida albcians persisters,