Qwin 500 image system

Additional samples of lung tissue fixed for 24 h in 10% neutral buffered formalin were dehydrated with ethyl alcohol and embedded in paraffin. The lung sections (5 mm³) were then stained with hematoxylin and eosin (H&E) according to the method described by Bancroft and Gamble [36 ] and examined under a light microscope. The histopathological lesions of lung sections stained with H&E were evaluated via histomorphometric analysis using ImageJ 1.46r software (Leica Qwin 500 image system, Cambridge, UK) [37 ]. The mean area of alveoli space and mean area percentages of inflammation and hemorrhage were measured.

Image J software was used to do histomorphometric analysis (Leica Qwin 500 image system, Cambridge, UK) [45 ] to evaluate histopathological lesions in H&E-stained sections. The mean area percentage of inflammatory mononuclear cells, bronchiole width in micrometers (µm), and thickening of interalveolar septa (µm) were assessed. The mean area % for iNOS, NF-kB, and Ki-67 immunoexpression intensity was evaluated. Each rat was evaluated by selecting 10 non-overlapping fields with 400×, a total magnification of 400× at random. The data were represented as means ± SEM (n = 6)