Masson s trichrome stain kit

A total of 28 days post-surgery, all rats were euthanized with 20% CO₂ for histological analysis. All heart tissues were quickly excised and dehydrated in a 30% sucrose solution. The hearts were then embedded in Tissue-Tek^® O.C.T.™ Compound (Sakura Finetek USA, Inc.) and cut into 7-µm slices. Modified Masson's Trichrome Stain kit (Beijing Solarbio Science & Technology Co., Ltd.) was used to examine infarct size according to the manufacturer's instructions. A light microscope (Leica Microsystems GmbH) was used to capture images (x1 magnification). Following Masson's staining, myocardial fibers appear red, while collagen fibers appear blue. The proportion of the infarct area was quantified using Imaging Pro Plus software 6.0 (Media Cybernetics, Inc.) and calculated as follows: Infarct heart area (%)=(endocardial length + epicardial length of the infarcted area/endocardial length + epicardial length of whole left ventricle) x100%.

Liver tissues were fixed in 4% paraformaldehyde and embedded in paraffin. Paraffin sections (5 μm) were then dewaxed and hydrated. Subsequently, the sections were sequentially stained with hematoxylin and eosin using a hematoxylin-eosin (HE) staining kit (Beyotime). Masson’s trichrome staining was performed to assess liver fibrosis, and sections were stained with Masson’s trichrome stain kit (Solarbio) according to the manufacturer’s instructions. For TUNEL staining, the sections were incubated with Biotin-dUTP, followed by staining with Streptavidin-HRP using the Colorimetric TUNEL Apoptosis Assay Kit (Beyotime). DAB was used to stain cell nuclei. Finally, the sections were examined under an optical microscope.

The heart tissues were dehydrated with sucrose solution (30%). The tissues were then embedded with Tissue-Tek (Sakura Finetek USA, Inc., USA), and sliced (7 µm). Masson’s Trichrome Stain kit (Solarbio, China) was used to stain the infarction area. The Masson positive area was red and the collagen fibers were blue. The images were taken by a light microscope (Leica, Germany).

Paraffin-embedded sections were routinely dewaxed, stained with the Masson's Trichrome Stain kit (Beijing Solarbio Science & Technology Co., Ltd.), according to the manufacturer's protocols, and finally, myocardial infarct size was calculated using Image ProPlus 6.0 (Media Cybernetics, Inc.).