Introduction: EUS is superior to any other imaging modalities with respect to the spatial resolution, but of limited value in producing parenchymal perfusion images with the use of the contrast agents. Recent discoveries of ultrasound have shown intravenous perfluorocarbon-containing microbubbles such as Definity produce harmonic signals at a low acoustic pressure. These contrast agents may be suitable for EUS that produces low acoustic pressure. In the present study, we observed microcirculation of digestive organs in dogs after the injection of Definity by means of a newly developed EUS probe. Methods: Contrast-enhanced harmonic sonography was conducted in 12 beagle dogs under anesthesia. The abdomen was opened by midline incision. An experimental EUS probe with a convex array was attached to stomach, small intestine, gallbladder or pancreas. The images were analyzed by ALOKA ProSound SSD 6500. E-PHD mode that sensitively depicts signals from microbubbles by the combination of filtered fundamental components and 2nd harmonic components was used. Each organ was sequentially observed after intravenous infusion of different doses of Definity. In some experiments, a part of the pancreas was ablated by radiofrequency after observing normal pancreas, and 1 ml of saline was injected into gastric submucosa. Results: After the injection of Definity, signals from microbubbles appeared from the area around the main pancreatic duct and subsequently branched off in the pancreas. The signals were finally distributed homogenously in the pancreatic tissue 10 seconds later. After the radiofrequency ablation, the contrast-enhanced harmonic sonography visualized the ablated lesion as a hypovascular perfusion defect. After vessels appeared flowing into the gallbladder, the muscle layer and the mucosal layer were sequentially observed. In stomach and small intestine, signals from microbubbles appeared as perforating vessels in the proper muscle layer and transverse vessels in the submucosal layer. Subsequently, signals were distributed in the mucosal layer. In the saline-injected lesion of gastric wall, some stretched vessels flowing between serosal and luminal sides were visualized. Conclusion: Contrast-enhanced harmonic sonography with the use of EUS probe successfully visualized microcirculation in digestive organs and would be a convincing method for depiction of small lesions, comprehension of detailed structure and differential diagnosis of digestive diseases. Introduction: EUS is superior to any other imaging modalities with respect to the spatial resolution, but of limited value in producing parenchymal perfusion images with the use of the contrast agents. Recent discoveries of ultrasound have shown intravenous perfluorocarbon-containing microbubbles such as Definity produce harmonic signals at a low acoustic pressure. These contrast agents may be suitable for EUS that produces low acoustic pressure. In the present study, we observed microcirculation of digestive organs in dogs after the injection of Definity by means of a newly developed EUS probe. Methods: Contrast-enhanced harmonic sonography was conducted in 12 beagle dogs under anesthesia. The abdomen was opened by midline incision. An experimental EUS probe with a convex array was attached to stomach, small intestine, gallbladder or pancreas. The images were analyzed by ALOKA ProSound SSD 6500. E-PHD mode that sensitively depicts signals from microbubbles by the combination of filtered fundamental components and 2nd harmonic components was used. Each organ was sequentially observed after intravenous infusion of different doses of Definity. In some experiments, a part of the pancreas was ablated by radiofrequency after observing normal pancreas, and 1 ml of saline was injected into gastric submucosa. Results: After the injection of Definity, signals from microbubbles appeared from the area around the main pancreatic duct and subsequently branched off in the pancreas. The signals were finally distributed homogenously in the pancreatic tissue 10 seconds later. After the radiofrequency ablation, the contrast-enhanced harmonic sonography visualized the ablated lesion as a hypovascular perfusion defect. After vessels appeared flowing into the gallbladder, the muscle layer and the mucosal layer were sequentially observed. In stomach and small intestine, signals from microbubbles appeared as perforating vessels in the proper muscle layer and transverse vessels in the submucosal layer. Subsequently, signals were distributed in the mucosal layer. In the saline-injected lesion of gastric wall, some stretched vessels flowing between serosal and luminal sides were visualized. Conclusion: Contrast-enhanced harmonic sonography with the use of EUS probe successfully visualized microcirculation in digestive organs and would be a convincing method for depiction of small lesions, comprehension of detailed structure and differential diagnosis of digestive diseases.