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. 2020 Sep 30;13:9623–9629. doi: 10.2147/OTT.S263250

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© 2020 Makino et al.

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Immunohistochemical (IHC) staining of ICAM-1. Lung tissues from the largest longitudinal section in the right lobe of control and smoking mice were stained for ICAM-1. Anti-ICAM1 primary antibody diluted 1:100 was used for IHC staining. ICAM-1 expression was quantified by measuring the percentage area of expression (0–100%) using ImageJ. Briefly, five randomly chosen field images (0.02 × 0.01 inches) from the slide containing the maximum longitudinal section of the right lobe were acquired using a light microscope, and colors that were not of interest were removed via the replace mode. The adapted images were converted to grey scale and the area of expression was located by adjusting the threshold. The percentage area of expression is denoted by positive pixels on the labeled areas. (A) IHC staining of ICAM-1 in control mouse lung. (B) IHC staining of ICAM-1 in smoking mouse lung. (C) The ICAM-1 antibody tended to strongly stain the lungs from the smoking group (7.75 ± 3.27 vs 5.48 ± 2.1, P = 0.049).