The aim of the present study was to evaluate the needle method for pancreatic tissue fluid pressure measurements. Clinical evaluation was performed in 24 patients with chronic pancreatitis, comparing repeated pressure measurements via sonographically guided fine-needle puncture and intraoperative pressure measurements by direct puncture of pancreatic tissue and duct. In patients with chronic pancreatitis we found small week-to-week variations in sonographically guided percutaneous pressure measurements and good agreement between preoperative percutaneous pressure measurements and intraoperative pressure measurements via direct puncture. Furthermore, no significant difference was seen between pancreatic duct and tissue fluid pressure. The technical evaluation was performed by repeated pressure measurements in human pancreatic autopsy specimens and living rats in a pressure chamber at various external pressure levels. The basic calibration of the method evaluated by means of this pressure chamber study showed sufficient precision and accuracy of the needle technique for clinical and investigative purposes. In conclusion, our results suggest that pancreatic tissue fluid pressure can be reliably assessed by the needle technique.