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.
Aniline blue dye solution
Manufactured by Merck Group
Sourced in United States
Aniline blue dye solution is a laboratory reagent used for staining and visualization purposes. It is a water-soluble dye that produces a blue color when applied to certain biological samples. The solution can be used for staining nucleic acids, polysaccharides, and other cellular components in microscopy and histological applications.
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Lab products found in correlation
Wiegert s iron hematoxylin solution, by Merck Group (3 mentions)
Masson ponceau acid fuchsin staining solution, by Merck Group (3 mentions)
Dm4000b, by Leica (1 mentions)
Optical microscope, by Nikon (1 mentions)
Wiegert s iron haematoxylin solution, by Merck Group (1 mentions)
Optical microscope, by Leica (1 mentions)
6 protocols using aniline blue dye solution
1
Masson's Trichrome Staining of Cardiac Tissue
2
Histological Staining and Imaging of Callus
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The sections were routinely deparaffinized and rehydrated in accordance with the aforementioned procedure. Weigert’s hematoxylin was used to dye the cell nucleus for 5 min. After rinsing with distilled water three times, the sections were stained in Masson-Ponceau-acid fuchsin solution for 10 min. They were rinsed in 2% glacial acetic acid and then differentiated in phosphomolybdic acid for 4 min. The sections were directly stained with aniline blue dye solution (Sigma). After dehydrating with ethanol series, clearing with xylene, and mounting with neutral resins, digital images were captured using an optical microscope (Leica DM 4000B) to analyze fiber formation in the early callus.
3
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).
4
Masson's Trichrome Staining of Myocardial Tissue
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After mice were anesthetized with i.p. ketamine (80 mg/kg)/xylazine (8 mg/kg) and sacrificed, the hearts were quickly excised, rinsed in cold PBS buffer, fixed in 40 μg/L paraformaldehyde for 72 h, dehydrated through ethanol, xylene, and cut into 5 μM thick sections. Nuclei were stained using wiegert’s iron haematoxylin solution for 5 min. After sections were rinsed three times with distilled water, 0.7% Masson-Ponceau-acid-fuchsin (Sigma-Aldrich, St. Louis, MO, USA) staining solution was added dropwise for 10 min, followed by 2% glacial acetic acid and phosphomolybdic acid dropwise for washing and differentiation, respectively. A 2% aniline blue dye solution (Sigma-Aldrich, St. Louis, MO, USA) was used for the final staining. Masson staining images and observation of myocardial tissue collagen were taken under a light microscope (Leica Microsystems, Wetzlar, Germany).
5
Histological Staining of Cardiac Tissue
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Mice cardiac tissues were fixed with 10% formaldehyde for 24 hours at room temperature, then decalcified, dehydrated, permeabilized with xylene, embedded in wax and finally sliced into 5‐μm‐thick sections with a microtome. Wiegert's iron haematoxylin solution (Sigma‐Aldrich, St. Louis, MO, USA) was used to dye the cell nucleus for 5 minutes. Following rinsing with distilled water 3 times, the sections were stained with 0.7% Masson‐Ponceau‐acid fuchsin staining solution (Sigma‐Aldrich) for 10 minutes. Samples were then 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.
6
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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