Spine is a structure commonly involved in several prevalent diseases. In clinical diagnosis, therapy, and surgical intervention, the identification and segmentation of the vertebral bodies are crucial steps. However, automatic and detailed segmentation of vertebrae is a challenging task, especially due to the proximity of the vertebrae to the corresponding ribs and other structures such as blood vessels. In this study, to overcome these problems, a probabilistic atlas of the spine, including cervical, thoracic and lumbar vertebrae has been built to introduce anatomical knowledge in the segmentation process, aiming to deal with overlapping gray levels and the proximity to other structures. From a set of 3D images manually segmented by a physician (training data), a 3D volume indicating the probability of each voxel of belonging to the spine has been developed, being necessary the generation of a probability map and its deformation to adapt to each patient. To validate the improvement of the segmentation using the atlas developed in the testing data, we computed the Hausdorff distance between the manually-segmented ground truth and an automatic segmentation and also between the ground truth and the automatic segmentation refined with the atlas. The results are promising, obtaining a higher improvement especially in the thoracic region, where the ribs can be found and appropriately eliminated.