This study aimed to assess the feasibility and safety of robotic-assisted navigation system for percutaneous transthoracic needle biopsy (PTNB), compare it with conventional freehand technique, and evaluate its generalizability across operators with varying experience levels. After excluding 5 patients in whom robotic-assisted PTNB could not be performed due to technical problems, a total of 50 patients with robotic-assisted PTNB and 200 patients who performed freehand puncture were included. Using propensity score matching (PSM) to match two groups of patients and simulate a randomized controlled scenario. The results showed that robotic-assisted PTNB significantly reduced the number of punctures, CT scans, and total procedure time (P <
0.05). These reductions were accompanied by a significantly lower rate of pneumothorax (P = 0.05), a common complication in PTNB procedures. While the overall adverse event rates remained similar between the two groups, the robotic-assisted technique demonstrated a more favorable safety profile, particularly with regard to reduced pneumothorax and hemorrhage rates. Additionally, there were no significant differences in the number of punctures, CT scans, total procedure time, and radiation dose administered to patients during robotic-assisted PTNB, irrespective of the operator. This suggests that operator experience does not significantly influence the outcomes of robotic-assisted PTNB, further highlighting the potential of the robotic system to minimize the impact of operator variability. Thus, we think robotic-assisted PTNB is feasible, safe, and less dependent on operator experience, suggesting its potential for clinical promote.