Endobronchial ultrasound (EBUS) improves bronchoscopic diagnosis of peripheral lung lesions (PLLs). The procedure time is lengthened by the search through multiple bronchial branches to PLLs. Virtual bronchoscopy (VB) provides endobronchial views that simulate findings at bronchoscopy. An open source software can be employed to study VB and hence the endobronchial route to a PLL. It allows VB to be studied in a Macintosh platform such as a laptop computer. Our purpose was to test if VB generated by this software could shorten procedure time of EBUS-guided bronchoscopy as compared with no VB assistance. The most feasible route to a PLL was manually selected using this software to study VB (VB group). For non-VB group, 2D CT axial +/- coronal images were reviewed to plan bronchoscopy. Before bronchoscopic biopsies, the location of PLL was confirmed by EBUS. Thirty-three subjects were recruited including 16 in VB group and 17 in non-VB group. The mean EBUS examination time and mean total procedure time were reduced in the VB group compared with non-VB group: 5.3 versus 10.5 minutes (P=0.04) and 22.4 versus 29.9 minutes (P=0.044), respectively. There was no complication in the VB group. Although the diagnostic yield was higher in the VB group than non-VB group, our study was not powered to test the difference. This pilot study suggests that VB assistance is safe and shortens procedural time of EBUS-guided bronchoscopy for PLL. Further study is warranted to confirm these findings.