Background/Objectives: Percutaneous transthoracic needle biopsy (PTNB) using advanced navigation techniques is increasingly performed; however, pulmonologists’ experience remains limited. This study reports an interventional pulmonologist’s initial experience with cone-beam computed tomography (CBCT) laser-guided PTNB and the diagnostic performance for lesions with diameters greater than or less than 20 mm. Methods: We retrospectively analysed the data of patients who underwent PTNB in a C-arm CBCT-equipped hybrid operating room between July 2020 and March 2024. All patients underwent the biopsy procedure under local anaesthesia. This was preceded by an initial 3D scan for planning of the needle route, followed by coaxial needle insertion. A post-procedural scan was also performed to identify complications. Results: Seventy-seven patients were enrolled in the study. The median distances of the needle path from the skin to the pleura and from the pleura to the lesion were 33.4 mm and 31.7 mm, respectively. The median number of tissue samplings was 4.9 ± 1.8. The median operating room duration was 51.5 ± 25.7 min, respectively. The median total dose area product was 8485.4 ± 5819.9 µGym². The sensitivity and specificity of our study findings were 93.3% (56/60) and 100%, while the accuracy was 94.8% (73/77). The overall complication rate was 13%. Conclusions: PTNB procedure by pulmonologists is a feasible and safe, single-operator workflow in a hybrid operating room. It can be performed under CBCT laser guidance with a similar diagnostic yield, acceptable radiation exposure and procedure duration, and minimal or manageable complications.