The aim of this study was to prospectively evaluate the needle visualization and placement error and use of an electromagnetic field-based tracking navigation device for puncture procedures based on C-arm CT (CACT) images. A commercially available navigation device was mounted on an angiographic X-ray system setup for CACT. After the target was defined, needle placement was performed under real-time visualization of the virtual needle in CACT images. The final, real needle position was assessed by CACT. Punctures were performed in phantoms (n = 76) and in twelve patients (eight biopsies, three drainages, one injection). Procedure times, system error, user error and total error were assessed. In phantoms, mean total error was 2.3 +/- 0.9 mm, user error was 1.4 +/- 0.8 mm and system error was 1.7 +/- 0.8 mm. In the patient study, the targeted puncture was successful in all twelve cases. The mean total error was 5.4 mm +/- 1.9 mm (maximum 8.1 mm), user error was 3.7 +/- 1.7 mm, system error was 3.2 +/- 1.4 mm and mean skin-to-target time was less than 1 min. The navigation device relying on CACT was accurate in terms of needle visualization and useful for needle placement under both experimental and clinical conditions. For more complex procedures, electromagnetic field-based tracking guidance might be of help in facilitating the puncture and reducing both the puncture risk and procedure time.