Background: Long linear lesions are created in the left atrium to modify the atrial substrate, thereby curing atrial fibrillation. The creation of long linear left atrial lesions using radiofrequency (RF) ablation is time consuming and difficult. Furthermore, it might result in significant complications. Cryoablation might overcome some of the disadvantages of RF ablation. Therefore, the aim of our study was to assess whether the creation of a long linear lesion is possible using cryotherapy. Methods: A right atrial septal linear lesion was created in six pigs (median weight: 50 kg; range: 40-60 kg). The ablation procedure was performed with a 7-F Freezor cryocatheter. The nonfluoroscopic mapping system LocaLisa was used as a navigation tool. At each point, freezing was maintained at the lowest attainable temperature (-75°C) for 4 minutes. The CARTO system was used for the evaluation of the linear lesions. Furthermore, all animals were sacrificed immediately after the ablation procedure and a postmortem examination of the lesions was performed. Additionally, an analysis of the amplitudes of the intracardiac electrograms registered via the ablation catheter was performed before and after the ablation procedure. Results: A right atrial septal linear lesion could be created successfully in all six pigs. For the performance of this ablation line, a median number of 16 cryoapplications (range, 11-26) was necessary. The amplitudes of the intracardiac electrograms registered via the ablation catheter decreased significantly after ablation. The CARTO bipolar voltage map revealed very low potentials along the ablation line and showed a sharply demarcated ablation area at the septum in all pigs. Further analysis of the CARTO map revealed an incomplete conduction block in all cases. Most of the pigs had a small gap close to the fossa ovalis. The postmortem examination of 2,3,5-triphenyl-tetrazolium chloride-stained specimens showed sharply demarcated lesions without any ulcerations. There were no major complications during the procedure. Conclusions: The creation of long linear lesions using cryoablation is feasible and safe. Lesion characteristics are different and more favorable than those created by RF. However, the aim of creating a transmural lesion and a complete conduction block remains an unsolved problem even with current cryoablation techniques. Nevertheless, growing experience and technical improvements might overcome some of the current limitations of this new technique.