Image-guided surgery can be broken down into two broad categories: frame-based guidance and frameless guidance. In order to reduce both the invasive nature of stereotactic guidance and the cost in equipment and time, we have developed a new guidance technique based on rapid prototyping (RP) technology. This new system first builds a computer model of the patient anatomy and then fabricates a physical reference frame that provides a precise and unique fit to the patient anatomy. This frame incorporates a means of guiding the surgeon along a preplanned surgical trajectory. This process involves (1) obtaining a high-resolution CT or MR scan, (2) building a computer model of the region of interest, (3) developing a surgical plan and physical guide, (4) designing a frame with a unique fit to the patient's anatomy with a physical linkage to the surgical guide, and (5) fabricating the frame using an RP unit. Software was developed to support these processes. To test the accuracy of this process, we first scanned and reproduced a plastic phantom fabricated to validate the system's ability to build an accurate virtual model. A target on the phantom was then identified, a surgical approach planned, a surgical guide designed, and the accuracy and precision of guiding a probe to that target were determined. Steps 1 through 5 were also evaluated using a head phantom. The results show that the RP technology can replicate an object from CT scans with submillimeter resolution. The fabricated reference frames, when positioned on the surface of the phantom and used to guide a surgical probe, can position the probe tip with an accuracy of 1.7 mm at the probe tip. These results demonstrate that the RP technology can be used for the fabrication of customized positioning frames for use in image-guided surgery.