Pancreatic cancer may present on computed tomography (CT) as an isolated cuff of tumor surrounding the superior mesenteric artery (SMA) or celiac trunk, without an identifiable pancreatic mass. We reviewed our experience with imaging-guided biopsy of the soft tissue cuff in this patient group. A retrospective review of our interventional database identified 163 patients referred for biopsy of suspected pancreatic carcinoma. Of these, eight patients underwent biopsy of an isolated cuff of soft tissue encasing the SMA (n = 6) or celiac trunk (n = 2). None of these eight patients had an identifiable pancreatic mass. The mean width of tissue cuff biopsied was 1.3 cm (range, 0.9-2.0 cm). Nine imaging-guided biopsies were performed in eight patients. Five biopsies were performed with color Doppler ultrasound and four with CT fluoroscopy. There was a median of two needle passes per procedure (range, 1-4). In six cases, a diagnosis of pancreatic adenocarcinoma was made at the first biopsy session. In one patient, ultrasound-guided biopsy was negative, but subsequent CT-guided biopsy was positive. In one additional patient with chronic pancreatitis, biopsy revealed benign fibrous tissue. There were no procedure-related complications. In patients with suspected pancreatic cancer (but without a focal parenchymal mass), imaging-guided biopsy of isolated periarterial tissue cuffs of tumor is accurate and safe.