To evaluate the efficacy of aspiration in an opposite position to deal with pneumothorax after CT-guided lung biopsy. A retrospective study was developed involving 210 patients with pneumothorax who had undergone CT-guided percutaneous core biopsies from January 2012 to March 2014 for various pulmonary lesions. Asymptomatic patients with minimal pneumothorax were treated conservatively. Simple manual aspiration was performed for symptomatic patients with minimal pneumothorax and for all patients with moderate to large pneumothorax. An opposite position aspiration was performed when simple manual aspiration failed. The efficacy of simple manual aspiration and the opposite position aspiration was observed. Among 210 patients with pneumothorax, 128 (61.0%) asymptomatic patients with minimal pneumothorax were treated conservatively. The remaining 82 were treated with attempted simple manual aspiration. Out of these 82 patients, simple manual aspiration was successful in 58 (70.7%, 58/82) cases. The complete and partial regression rates were 17.2% (10/58) and 82.8% (48/58), respectively. In the other 24 patients (29.3%, 24/82), simple aspiration technique was ineffective. An opposite position (from prone to supine or vice versa) was applied, and a new biopsy puncture site was chosen for reaspiration. This procedure was successful in 22 patients but not in 2 patients who had to have a chest tube insertion. The complete and partial regression rates were 25.0% (6/24) and 66.7% (16/24), respectively. Applying the new method, the total effective rate of aspiration improved significantly from 70.7% (58/82) to 97.6% (80/82). The opposite position aspiration can be safe, effective and minimally invasive treatment for CT-guided lung biopsy-induced pneumothorax thus reducing the use of chest tube significantly. (1) Opposite position aspiration can elevate the success rate of aspiration significantly (from 70.7% to 97.6% in our study); (2) this procedure is a safe, effective and minimally invasive treatment for pneumothorax caused by biopsy; and (3) opposite position aspiration is a useful technique to reduce the use of chest tube, which has clinical significance.