Percutaneous transluminal biopsy performed under fluoroscopic control uses interventional access routes and allows direct sampling of biliary tract lesions. The authors report their experience with this technique in 20 patients. The biopsy site was the gallbladder in 9 patients and the intrahepatic or extrahepatic bile ducts in 11. A suspected malignant lesion was the indication for biopsy in 17 patients; 3 patients underwent biopsy of the gallbladder mucosa during ablation therapy. The procedure was performed with the forceps (for 17 patients), brush (for 2) or "scrape" (for 1) technique; diagnostic material was obtained in all but one case. The sensitivity, specificity and overall accuracy of diagnosis in the patients with a suspected malignant lesion were 71%, 100% and 88% respectively. There were no false-positive results. In this series one major complication, hemobilia necessitating transarterial embolization, occurred after the scrape biopsy, and minor, self-limiting gallbladder hemorrhage occurred in one patient. This study shows that percutaneous transluminal biopsy is a reliable technique for examining a variety of biliary tract lesions and can lead to an accurate pathological diagnosis.