We have used a two-step casting procedure to create replicas of fibrillar collagen. A negative replica or mold is first created using either Mercox casting resin or a prepolymerized methacrylate resin that was originally described by Murakami. In the second step, a positive replica is made from the first using a solution of polystyrene or polyurethane to coat the methacrylate cast. The resulting replica is removed from the methacrylate cast. Results indicate the highest level of spatial resolution is obtained using a modified Murakami resin for the negative cast and polystyrene for the positive replica. The 29-nm-wide repeat banding structure present on collagen fibrils could be clearly identified in both the methacrylate cast and in the polystyrene positive replica of the methacrylate cast. Thus high-resolution replicas can be obtained. Such replicas may be useful to imprint inplantable devices with biologically relevant textures to improve tissue integration or for the development of textured surfaces for the in vitro expansion of cultured cells.