Fig. 7.

CX₃CR1 CD14 immunofluorescent staining of the pulmonary microvasculature of COPD, S and NS. These figures are representative images for dual label immunofluorescent detection of CD14 by CX₃CR1⁺ monocytes marginated within pulmonary microvessels in human lung tissue. Representative images 6a-c) n = 9 COPD, 6d-f) n = 9 S and 6 g-i) n = 6 NS. Cell nuclei were counterstained with 4’,6-diamidino-2-phenylindole (blue). CX₃CR1⁺ cells were identified using an Alexa-568 conjugated donkey anti-rabbit secondary antibody (red 6a, 6d, 6 g). CD14⁺ cells were labelled with a biotinylated rabbit anti-goat secondary antibody and detected using Streptavidin Dylight 488 (green 6b, 6e, 6 h). Composite images are shown (6c, 6f, 6i). Green/yellow autofluorescence is caused by intrinsically fluorescent tissue components such as elastic fibres and erythrocytes. Autofluorescence can be distinguished from positive fluorescence by forming a composite image of the red, green and blue channels. Autofluorescence is visible in all three channels and so appears as an amalgamation of the three colours. Positive fluorescence is visible in one channel only and thus appears as the pure colour. Singly labelled cells appear in the composite image as either being red or green, dual labelled cells appear yellow. Examples of immunoreactive cells are indicated by arrows (images taken using X20 objective lens). The white scale bar represents a length of 75 μm