BACKGROUND: Conventional sputum suctioning is a routine clinical practice, but complications may arise from the blind manipulation of the catheter. Recently, a visual sputum suctioning system (VSSS) was developed, and we tested this new system in a laboratory setting. METHODS: We used coagulant concentrations of 1.5% and 3.0% to simulate mucus and sputum. Conventional singlelumen and triple-lumen catheters were inserted separately into a beaker for sputum suctioning (15 s, 200 mm Hg). A micro-imaging fiber was integrated into the triple-lumen catheter to create the VSSS. The single-lumen catheter and the VSSS were inserted separately into the mouth cavity, the nasal cavity, the tracheostomy tube, and the endotracheal tube of a human analog model for further comparisons. RESULTS: As the suction channel of the triple-lumen catheter was reduced by 46.8%, the amount of simulant it suctioned was significantly less than that suctioned by the single-lumen catheter. However, under real-time guidance, the VSSS suctioned more simulant than the conventional single-lumen catheter in the human analog model. CONCLUSION: Sputum suctioning with the VSSS was feasible. Because of its real-time imaging guidance, the efficiency of the VSSS procedure was greater than that of the conventional single-lumen catheter. Therefore, this system may provide a new platform for sputum suctioning.