Analyzing Macrophage Infiltration in Aortic Plaques

Mice were euthanized (n = 3–4/group), perfused with PBS (5 mL) and the aortic arch with heart tissue was embedded in OCT freezing compound. Seven-micrometer serial cryosections were collected every 70 µm in the innominate artery. Immunohistochemistry was performed on cryosections corresponding to greatest plaque accumulation in the innominate artery as previously described [42] (link), [53] (link) using rat anti-mouse F4/80 (no. MCA497R; Serotec, Oxford, U.K.) or isotype controls (no. MCA1125; Serotec). Biotinylated anti-rat (mouse absorbed) IgG was used as secondary Ab (Vector Laboratories, Burlin- game, CA). Nuclei were counter-stained with hematoxylin. Digital micrographs were captured at 10× and 40×.