Computational protocol: Disrupting the Btk Pathway Suppresses COPD-Like Lung Alterations in Atherosclerosis Prone ApoE−/− Mice Following Regular Exposure to Cigarette Smoke

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Protocol publication

[…] For histochemical analysis, fixed lungs were embedded in paraffin and sectioned for staining. Tissue sections were stained with Hart’s Elastin, Picro-Sirius Red (PSR), Hematoxylin and Eosin.Differences in hue of PSR red stained airways were evaluated using ImageJ software. For airway analysis images were converted to RGB format, then the airway was selected as the region of interest (ROI) and mean intensities of only the red components were measured to exclude background and thin (green) fibers. Results were expressed as red intensity for each ROI. Differences in lung vasculature collagen content were measured in ImageJ by converting color images to gray-scale, selecting vasculature as the ROI, and measuring mean intensities. MLI was calculated to quantify airspace enlargement according to a previously described method [] using Hematoxylin and Eosin stained lung tissue sections. Hart’s Elastin stained lung sections were employed to analyze a destruction of alveolar walls (breaks in elastic fibers) [,,,]. [...] Results are expressed as the mean ± STD. Differences between multiple groups were evaluated by one way ANOVA or the non-parametric Kruskal-Wallis ANOVA on ranks as dictated by variance/normality followed by post hoc analysis with Fisher’s least significant difference test or Dunn’s multiple comparison test respectively. In cases where Dunn’s test was unable to yield a direct evaluation for significance comparisons between groups of interest were performed using the nonparametric Mann-Whitney test with Bonferroni correction for multiple comparisons. Differences between paired groups were evaluated with Student’s t-test or the non-parametric Mann-Whitney rank sum test as dictated by variance/normality. The specific analyses used are indicated in individual figure legends. All statistics were performed using SIGMAPLOT 11 (Systat Software Inc., San Jose, CA, USA). Significance was defined as p < 0.05. […]

Pipeline specifications

Software tools ImageJ, SigmaPlot
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Mus musculus, Homo sapiens
Diseases Pulmonary Disease, Chronic Obstructive, Atherosclerosis
Chemicals Tyrosine