An accurate and accessible image segmentation method is in high demand for generating 3D bone models from CT scan data, as such models are required in many areas of medical research. Even though numerous sophisticated segmentation methods have been published over the years, most of them are not readily available to the general research community. Therefore, this study aimed to quantify the accuracy of three popular image segmentation methods, two implementations of intensity thresholding and Canny edge detection, for generating 3D models of long bones. In order to reduce user dependent errors associated with visually selecting a threshold value, we present a new approach of selecting an appropriate threshold value based on the Canny filter. A mechanical contact scanner in conjunction with a microCT scanner was utilised to generate the reference models for validating the 3D bone models generated from CT data of five intact ovine hind limbs. When the overall accuracy of the bone model is considered, the three investigated segmentation methods generated comparable results with mean errors in the range of 0.18–0.24 mm. However, for the bone diaphysis, Canny edge detection and Canny filter based thresholding generated 3D models with a significantly higher accuracy compared to those generated through visually selected thresholds. This study demonstrates that 3D models with sub-voxel accuracy can be generated utilising relatively simple segmentation methods that are available to the general research community.