Figure 4
From: Activated neutrophil fluorescent imaging technique for human lungs
Representative OEM images from participants with bronchiectasis before and after the application of NAP are shown in the left-hand image columns. One participant received NAP but did not tolerate subsequent imaging, so five sets of imaging are shown. Blue arrows—alveolar wall, red arrow—punctate fluorescent NAP signal, purple arrows—pulmonary capillary. The labelling of relevant features is not exhaustive in these representative images. Laboratory confocal microscopy from the broncho-alveolar lavage is shown with each pair of OEM images. Red stain—Calcein red cytoplasmic stain. Purple stain—Styo62 nuclear stain. Green stain—material excited at 488 nm and emitting at 500–530 nm. Calcein red and Syto62 are applied after lavage harvest. Green fluorescent staining has taken place within the lung. Left hand side: selected clinical parameters from each participant are shown. Right hand side: pooled subjective blinded video analysis from two blinded experienced observers working independently is shown. 1—no suspicion of NAP signal, 2—suspicion of NAP signal, 3—positive NAP signal, 4—very positive NAP signal. Of the CD66bhi population in lavage, values for CD11bhi and CD63hi cell surface markers as a ratio of the relevant isotype control are shown. Bottom row; larger example OEM images for inspection, description given with image. Scale bar OEM 50 µm, scale bar laboratory confocal microscopy 50 µm.
