Histological Analysis of Infarcted Tissue

Tissues were immersed in 4% paraformaldehyde for 4 h and transferred to 70% ethanol. Then, the tissues were placed in cassettes, dehydrated through a serial alcohol gradient, and embedded in paraffin wax blocks. Histopathological examination was performed in infarcted tissue on formalin-fixed, paraffin-embedded 5–6 μm sections stained with hematoxylin and eosin using standard methods and examined via light microscopy (Mak et al., 2020 (link)). Primary antibodies against CD68 (1:500, Abcam, #ab31630) and eNOS (1:300, Abcam, #ab76198) were used. A TUNEL assay kit was used according to the manufacturer’s instructions (R&D Systems). HE staining was used to detect the arrangement of RBC morphology to evaluate the integrity of microvessels.