Aiming devices enable the use of neuronavigation systems for rigid instrument guidance mimicking the possibilities of a frame-based system without having a stereotactic frame affixed to the skull. The aim of the presented work was to investigate the phantom targeting accuracy of the Vertek aiming device (Medtronic Inc., Louisville, USA) and whether it can be safely and accurately applied in a concept of minimally invasive brain biopsy in which multi-modal image fusion, image-to-patient registration and head immobilization were based on a non-invasive vacuum mouthpiece. A plastic model of a head with 20 target beads broadly distributed around the head volume was used for determination of CT-based targeting accuracy. Every target was punctured 5 times totaling 100 needle positionings. Accuracy was evaluated on postoperative CT scans with the needles in place. The mean normal deviation (n = 100) was 1.5 +/- 0.8 mm and the mean angle of deviation was 1.1 +/- 0.7 degrees. In a preliminary clinical series in ten patients diagnostic biopsy sampling of intracranial lesions with a median diameter of 28 mm (range: 12-90 mm) could be achieved in all patients and no biopsy related complications were recorded. The experimental results showed a similar accuracy to frame-based stereotaxy. The device facilitates trajectory alignment via two pivot joints and the actual depth and location of the biopsy needle can be monitored. Within the limitations of a preliminary study, brain biopsy may be accurately and safely performed for lesions > or = 12 mm.