Background and Study Aims: The accuracy of endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) depends on immediate specimen review by a cytopathologist. Stromal tumors, lymphoma, and well-differentiated pancreatic cancer are difficult to diagnose on the basis of cytology alone. To overcome these limitations, a 19-gauge Trucut needle has been developed to obtain histological samples at EUS. This pilot study compares the specimen adequacy and diagnostic accuracy of EUS-guided Trucut needle biopsy (EUS-TNB) with EUS-FNA. Patients and Methods: A total of 18 patients underwent EUS-TNB and EUS-FNA. The specimen adequacy and diagnostic accuracy of the two techniques was compared. The technical performance and safety profile of the Trucut needle were also evaluated. Results: The EUS-TNB specimen was adequate for evaluation in 15/18 patients compared with 18/18 with EUS-FNA (83 % vs. 100 %, not significant). The diagnostic accuracy of EUS-TNB was not significantly different from EUS-FNA (78 % vs. 89 %). Two complications were encountered: one patient developed mediastinitis and required surgery; another had immediate bleeding that was managed conservatively. One technical problem was encountered: the Trucut needle failed to deploy after two passes when a gastric stromal cell tumor was being biopsied. Conclusion: The diagnostic accuracy of the new EUS-TNB is comparable to that of EUS-FNA. In our experience, the overall efficacy and safety profile of the Trucut needle appears modest.