Objective To investigate the clinical value of empty needle navigation combined with quiet breathing in CT-guided puncture biopsy of lung subcentimeter nodules.Methods A retrospective analysis was conducted of sixty-three patients with subcentimeter pulmonary nodules (≤1 cm) treated in our hospital. Puncture materials were collected by using empty needle navigation combined with quiet breathing under the guidance of CT, and the success rate of puncture biopsy, postoperative complications and results of postoperative pathological diagnosis were statistically analyzed.Results The success rate of puncture biopsy was 98.4% (62/63). The incidence of pneumothorax and hemorrhage was 20.6% (13/63) and 31.7% (20/63)respectively. Neither infection nor tumor spread or metastasis were found during the follow-up. Among the 62 patients with successful biopsy, 4 cases of squamous cell carcinoma, 21 cases of adenocarcinoma (3 cases of adenocarcinoma in situ), 2 cases of metastatic carcinoma and 1 case of malignant melanoma were reported. There were 19 cases of chronic inflammation, 7 cases of granulomatous inflammation, 5 cases of dust accumulation accompanied by lymphocytic hyperplasia, 2 cases of alveolar epithelial hyperplasia, and 1 case of fungus.Conclusions Puncture biopsy in patients with subcentimeter pulmonary nodules by using empty needle navigation combined with quiet breathing is a safe and reliable method with a high success rate. It is an important means for early diagnosis of lung cancer.