The masson staining was conducted as previously described [19 ]. The cardiac tissues were firstly fixed with 10% formaldehyde for 24 hours at room temperature. After decalcified, dehydrated, permeabilized with xylene, the tissues were embedded in wax and finally cut into 5‐μm‐thick sections. The cell nucleus was dyed for 5 minutes using the Wiegert’s iron hematoxylin solution (Sigma Aldrich, St. Louis, MO, USA). After rinsed with distilled water for at least 3 times, the sections were stained with 0.7% Masson-Ponceau-acid fuchsin staining solution (Sigma-Aldrich) for 10 minutes at room temperature. Then, the sections were rinsed in 2% glacial acetic acid and differentiated in phosphomolybdic acid for 4 minutes. The sections were directly stained with 2% aniline blue dye solution (Sigma-Aldrich). Following dehydrating with ethanol series, clearing with xylene and mounting with neutral resins, images of the stained sections were captured with a light microscope.
Wiegert s iron hematoxylin solution
Manufactured by Merck Group
Sourced in United States
Wiegert's iron hematoxylin solution is a laboratory reagent used in histological and cytological staining procedures. It is a complex mixture of hematoxylin, ferric salts, and other ingredients that produces a dark blue-black stain, commonly used to visualize nuclei in tissue sections or cell preparations. The solution provides a reliable and consistent staining outcome for microscopic analysis.
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3 protocols using wiegert s iron hematoxylin solution
1
Masson's Trichrome Staining of Cardiac Tissue
2
Fetal Heart Tissue Histological Analysis
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Fetal heart tissue was fixed with 10% FA at room temperature for 24 h, then decalcified, dehydrated, permeated with xylene, embedded in wax. Wiegert's iron hematoxylin solution (Sigma-Aldrich) was used to stain the nuclei for 5 min. First stain with 0.7% Masson's Trichrome Stain solution (Sigma-Aldrich) for 10 min after rinsing with distilled water. Rinsing: 2% glacial acetic acid; differentiation: in phosphomolybdic acid, 4 min. Continue dyeing with 2% aniline blue dye solution (Sigma-Aldrich). After dehydration, dewaxing, and fixation with neutral resin, the image was taken with an optical microscope (Nikon).
3
Histopathological Analysis of Myocardial Tissue
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The histopathological examination of myocardial tissue was stained with hematoxylin-eosin (HE). The left ventricle of the heart sample was placed in a 10% formaldehyde solution, dehydrated with ethanol gradient, embedded in paraffin, and cut into 4 μm sections. After the sample was removed, it was stained with hematoxylin and eosin. Then, the slices were observed under an optical microscope (Leica Microsystems, Wetzlar, Germany). For Masson staining, the rat heart tissue sections were firstly used to stain cell nuclei for 5 min by Wiegert's iron hematoxylin solution (Sigma-Aldrich, St. Louis, MO, USA). After rinsing 3 times with distilled water, the sections were stained with 0.7% Masson-Ponceau-acid fuchsin staining solution (Sigma-Aldrich) for 10 min. After rinsing in 2% glacial acetic acid and differentiated in phosphomolybdic acid for 4 min, the sections were directly stained with 2% aniline blue dye solution (Sigma-Aldrich). After dehydration with ethanol series, remove with xylene, install with neutral resin, and capture the image of the stained section with an optical microscope
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