Initial histopathological analysis of a pulmonary lesion is mandatory whenever a lung cancer is suspected in order to determine the optimal diagnostic strategy. Adequate material must be obtained for a definite histological diagnosis (preferable to a cytological one) and in some cases (mainly in adenocarcinoma) molecular analysis. The two main methods to obtain adequate biopsy samples are flexible bronchoscopy and computed tomography (CT)-guided lung biopsy. Flexible bronchoscopy is a valuable method for proximal tumors. On the other hand, CT-guided lung biopsy is more accurate for peripheral tumors. CT-guided lung biopsy is a reliable procedure that conveys a 90% sensitivity for the diagnosis of lung cancer. When performed in a secured environment after contraindications evaluation, its severe complications rate is low, mainly consisting of pneumothorax requiring chest tube placement and hemoptysis.