Transarticular C1/2 screws are widely used in posterior cervical spine instrumentation. The use of pedicle screws in the cervical spine remains uncommon. Due to superior biomechanical stability compared to lateral mass screws, pedicle screws can be used, especially for patients with poor bone quality or defects in the anterior column. Nevertheless there are potential risks of iatrogenic damage to the spinal cord, nerve roots or the vertebral artery associated with both posterior cervical spine instrumentation techniques. Therefore, the aim of this study was to evaluate whether C1/2 transarticular screws as well as transpedicular screws in C3 and C4 can be applied safely and with high accuracy using a computer-assisted surgery (CAS) system. We used 13 human cadaver C0-C5 spine segments. We installed 1.4-mm Kirschner wires transarticular in C 1/2, using a specially designed guide, and drilled 2.5-mm pedicle holes in C3 and C4 with the assistance of the CAS system. Hole positions were evaluated by palpation, CT and dissection. Forty-eight (92%) of the 52 drilled pedicles were correctly positioned after palpation, imaging and dissection. The vertebral artery was not injured in any specimen. All of the 26 C1/2 Kirschner wires were placed properly after imaging and dissection evaluations. No injury to vascular or bony structures was observed. C /2 transarticular screws as well as transpedicular screws in the cervical spine can be applied safely and with high accuracy using a CAS system in vitro. Therefore, this technique may be used in a clinical setting, as it offers improved accuracy and reduced radiation dose for the patient and the medical staff. Nevertheless, users should take note of known sources of possible faults causing inaccuracies in order to prevent iatrogenic damage. Small pedicles, with a diameter of less than 4.0 mm, may not be suitable for pedicle screws.