Background: Obtaining tissue from a para-aortal lymph node or tumor is a challenge that Currently requires invasive Surgical procedures. Para-aortic lung tumors can be clearly visualized by EUS. Although the accessibility of lesions adjacent to the esophagus is well documented, the para-aortic region has never been systematically explored. Objective: To assess the feasibility, yield, and safety of transaortic biopsy specimens in the diagnosis of lung Minors and nodal masses located lateral to the aorta. Design: A retrospective case series of 14 consecutive patients. Setting: Pulmonary Department, Leiden University Medical Center, Leiden, The Netherlands. Patients: Fourteen patients with known or Suspected lung cancer. Nine patients presented with a left-sided lung mass (mean size 27 mm), whereas 5 patients had an enlarged para-aortic node (mean size 16 mm). Interventions: Real-time EUS-guided transaortic biopsy of a para-aortic lesion. Main outcome measurements: Feasibility, diagnostic yield, and complication rates of transaortic EUS-guided FNA (EUS-FNA). Results: The final diagnosis was known in 12 patients (10 non-small-cell lung carcinoma [NSCLC], 1 small-cell lung carcinoma [SCLC], and 1 renal-cell carcinoma). EUS-FNA established malignancy in 9 of 14 patients (64%) (8 NSCLC and 1 SCLC). One aspirate revealed reactive nodal tissue, and 4 demonstrated nonrepresentative material. Malignancy was further assessed in 3 patients after subsequent diagnostics. Transaortic FNA was found to be safe. In 2 patients, EUS images after biopsy were suspicious for a small para-aortic hematoma. These patients recovered uneventfully Conclusions: These results demonstrate that a single EUS-guided transaortic biopsy of para-aortic lymph nodes and tumors is a feasible and probably safe method that results in a diagnosis in the majority of cases.