Referring to prior research [32 (link)], following fixation with 4% paraformaldehyde and paraffin-embedding, the cardiac tissues were sliced into sections (thickness of 5 μm) and cultured with hematoxylin and eosin solution (H&E, Sigma, St Louis, MO, USA) and Masson’s trichrome (Polysciences, Bay Shore, N.Y., USA) for observation of the myocardial histopathological structure and myocardial fibrosis.
All cardiac sections were collected and stored digitally using a Nikon slide scanner. To evaluate myocardial fibrosis, 20X Massons-Trichrome images of the entire LV were exported, and the Image-J software (National Institutes of Health, Bethesda, MD, USA) was adopted to calculate the collagen fraction (%) as the ratio of the total fibrosis area (blue) to the total tissue area. The above-mentioned experiments were performed by independent technicians who were blinded to mice grouping.
All cardiac sections were collected and stored digitally using a Nikon slide scanner. To evaluate myocardial fibrosis, 20X Massons-Trichrome images of the entire LV were exported, and the Image-J software (National Institutes of Health, Bethesda, MD, USA) was adopted to calculate the collagen fraction (%) as the ratio of the total fibrosis area (blue) to the total tissue area. The above-mentioned experiments were performed by independent technicians who were blinded to mice grouping.
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