Figure 5
NBD-UBIdend labels colonised bacteria in situ in whole explant Cystic Fibrosis (CF) Human Lungs. (A) Experimental procedure of CF lungs with bronchoscopy, Smartprobe instillation, OEM imaging and bronchoalveolar lavage retrieval. (B) Representative frames of segments instilled with NBD-UBI (6 µM) with minimal punctate signal (upper) and segment instilled with NBD-UBIdend (2 µM) demonstrating the characteristic punctate signal associated with bacterial labelling. (C) Analysis of OEM videos with image analysis algorithm showing a significantly higher proportion of frames containing >80 dots per frame (bacterial infection). Bars represents the combined mean (+/− SEM) of the percentage of frames with over 80 detected dots per frame over the imaging sequence for individual segments, **p < 0.01, n = 3 CF patient explants. (D) Lavage counts from BALF retrieval for control, n = 3 from two CF lungs and NBD-UBIdend, n = 4 from three CF lungs. Bars represent mean (+/− SD) of counts.
