In the era of lung cancer screening and early detection of lung lesions, pulmonary segmentectomy has gained wide acceptance between thoracic community reducing the need of lobectomy for diagnostic purpose and treatment in case of centrally located benign, multiple or undetermined lesions. In rigorously selected patients with stage I non-small cell lung cancer (NSCLC), segmentectomies seem to offer similar survival outcomes rather than lobectomies, but associated with a better conservation of lung function. However, segmentectomy is a more challenging procedure to be performed compared to lobectomy, especially by video-assisted thoracic surgery (VATS). Many difficulties could arise during video-assisted segmentectomy, making the procedure more demanding and stressful. Following the introduction of the near infrared (NIR)/indocyanine imaging system on standard endoscopic module, we decided to adopt peripheral intravenous injection of indocyanine green (ICG) to identify intersegmental plain during uniportal VATS lung segmentectomy. Our technique herein is widely illustrated.