Characteristics of the swelling-induced wrinkles on the surfaces of natural rubber (NR) film were investigated. The wrinkle structure was generated by swelling of NR film pre-stretched and firmly bonded onto an aluminum substrate in hexane. A novel experimental method was adopted to replicate the swelling-induced wrinkles on the NR film using an epoxy-hardener system. To get insight into the wrinkle parameters; the wrinkle length (L), wrinkle distance (D), wrinkle height (H) and the angle between two consecutive wrinkles (θ), the cross-sections of the replicas obtained from saturated swollen NR film were examined using an optical microscopy (OM). From the OM images, the wrinkling parameters were measured as a function of the thickness of NR film from 0.42 to 1.76 mm. Also, it was evaluated that the effects of swelling time on the wrinkling parameters. The length (L), distance (D) and height (H) of wrinkles increased as the thickness of the NR film and the swelling time increased. However, the angle between the wrinkles (θ) showed a sharp decrease up to a swelling time of 200 minutes and slightly decreased afterwards.