The objective of the present study was to assess feasibility of single port (uniportal) VATS in the diagnosis and staging of mediastinal nodal enlargements. To this purpose, between January 2002 and October 2005, 13 patients (8 males and 5 females; mean age 54 years) have undergone uniportal VATS sampling of mediastinal nodes either as part of a diagnostic pathway or as a staging procedure for primary lung cancer when either nodal stations were inaccessible to standard mediastinoscopy or in the event of redo nodal biopsy. Sampled stations were the aortopulmonary window (6), subcarinal (1), right paratracheal (5), and, paraesophageal (1) ones. In all cases, sufficient samples were made available for pathological diagnosis. Postoperatively, neither morbidity nor mortality was observed. The median length of stay in the hospital was 1 day. In conclusion we showed that uniportal VATS can be effectively used to achieve diagnosis and staging of mediastinal nodal stations.