BACKGROUND: Ultrathin bronchoscopy guided by virtual bronchoscopy (VB) techniques show promise for the diagnosis of peripheral lung lesions. In a phantom study, we evaluated a new real-time, VB-based, image-guided system for guiding the bronchoscopic biopsy of peripheral lung lesions and compared its performance to that of standard bronchoscopy practice. METHODS: Twelve bronchoscopists of varying experience levels participated in the study. The task was to use an ultrathin bronchoscope and a biopsy forceps to localize 10 synthetically created lesions situated at varying airway depths. For route planning and guidance, the bronchoscopists employed either standard bronchoscopy practice or the real-time image-guided system. Outcome measures were biopsy site position error, which was defined as the distance from the forceps contact point to the ground-truth lesion boundary, and localization success, which was defined as a site identification having a biopsy site position error of < or = 5 mm. RESULTS: Mean (+/- SD) localization success more than doubled from 43 +/- 16% using standard practice to 94 +/- 7.9% using image guidance (p < 10(-15) [McNemar paired test]). The mean biopsy site position error dropped from 9.7 +/- 9.1 mm for standard practice to 2.2 +/- 2.3 mm for image guidance. For standard practice, localization success decreased from 56% for generation 3 to 4 lesions to 31% for generation 6 to 8 lesions and also decreased from 51% for lesions on a carina vs 23% for lesions situated away from a carina. These factors were far less pronounced when using image guidance, as follows: success for generation 3 to 4 lesions, 97%; success for generation 6 to 8 lesions, 91%; success for lesions on a carina, 98%; success for lesions away from a carina, 86%. Bronchoscopist experience did not significantly affect performance using the image-guided system. CONCLUSIONS: Real-time, VB-based image guidance can potentially far exceed standard bronchoscopy practice for enabling the bronchoscopic biopsy of peripheral lung lesions.