Alcian blue (AB) staining indicates the deposition of sulphated glycosaminoglycans in blue. For AB staining, the deparaffinized sections were incubated for 3 min in 1% acetic acid (Carl Roth GmbH and Co.KG) and then immersed for 30 min in 1% AB in acetic acid (Carl Roth GmbH and Co.KG). Subsequently, they were rinsed in 3% acetic acid. Cell nuclei were counterstained for 5 min with nuclear fast red aluminum sulphate solution (Carl Roth GmbH and Co.KG).
Nuclear fast red aluminum sulphate solution
Manufactured by Carl Roth
Sourced in Germany
Nuclear fast red aluminum sulphate solution is a laboratory dye used for staining nucleic acids in various analytical and research applications. It provides a reddish-pink color to DNA and RNA samples, enabling their visualization and identification during electrophoresis, microscopy, and other experimental procedures.
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Lab products found in correlation
Acetic acid, by Carl Roth (2 mentions)
Entellan, by Merck Group (2 mentions)
Dm1000 led, by Leica (2 mentions)
Roti histol, by Carl Roth (1 mentions)
Roti histokitt, by Carl Roth (1 mentions)
Histogel, by Thermo Fisher Scientific (1 mentions)
Xylol, by Carl Roth (1 mentions)
Harry s hematoxylin, by Carl Roth (1 mentions)
4 protocols using nuclear fast red aluminum sulphate solution
1
Alcian Blue Staining for Glycosaminoglycans
2
Alcian Blue Staining of Glycosaminoglycans
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Alcian blue (AB) staining indicates the deposition of sulphated glycosaminoglycans by a blue color. For AB staining, the deparaffinized sections were incubated for 3 min in 1% acetic acid (Carl Roth GmbH and Ko.KG) and then immersed for 30 min in 1% AB in acetic acid (pH 2.5) (Carl Roth GmbH and Ko.KG). Subsequently, they were rinsed in 3% acetic acid. Cell nuclei were counterstained using nuclear fast red aluminum sulphate solution (Carl Roth GmbH and Ko.KG) for 5 min. The sections were dehydrated in a series of ascending alcohol concentrations (70, 80, 96, and 99.6%) before being embedded in Entellan (Merck KGaA) and kept in the dark until analyzed by light microscopy (DM1000 LED, Leica Microsystems GmbH). AB scoring was performed according to the following criteria:
The staining intensity was slightly or substantially lower or higher as the native rabbit ACL (1 or 0 points) or the staining intensity was similar to the native rabbit ACL (2 points). The staining was homogeneously distributed (2 points) or slightly (1 point) or severely inhomogeneously stained (0 point). Hence, the possible maximum was 4 points.
The staining intensity was slightly or substantially lower or higher as the native rabbit ACL (1 or 0 points) or the staining intensity was similar to the native rabbit ACL (2 points). The staining was homogeneously distributed (2 points) or slightly (1 point) or severely inhomogeneously stained (0 point). Hence, the possible maximum was 4 points.
3
Prussian Blue Staining for Ferric Iron
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We analyzed ferric iron (Fe3+) by Perl’s Prussian blue staining [62 ]. A total of 30 µm sections of frozen adipose tissue were fixed with 4% buffered neutral formalin at room temperature for 5 min, washed three times with distilled water, and air-dried. After descending alcohol series, the tissue was pre-stained in 10% potassium-ferrocyanide for 5 min, and stained with equal parts of a mixture of 2% potassium-ferrocyanide solution and 1% hydrochloric acid for 30 min at 37 °C. After washing with distilled water, the slides were counterstained with nuclear fast red aluminum sulphate solution (Roth, Germany) for 5 min, rinsed with distilled water, dehydrated in Roti-Histol (Roth, Germany), and mounted in Roti-Histokitt (Carl Roth, Germany).
4
Histological Analysis of Cell Sheets
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The cell sheets were fixed for 15 min with 4% paraformaldehyde (PFA) and placed inside HistoGel (ThermoFisher Scientific Inc., Germany) before being embedded in paraffin. Sections of 7 µm were prepared. The sections were 10 min deparaffinized in xylol (Carl Roth GmbH and Ko.KG) and rehydrated with a descending ethanol row (ETOH, 99.8%, 96%, 80%, 70%) (Carl Roth GmbH and Ko.KG).
For hematoxylin-eosin (HE) staining, sections of sheets were incubated for 6 min in Harry’s hematoxylin (Carl Roth GmbH and Ko.KG), before being rinsed in running tap water and counterstained for 4 min in eosin (Carl Roth GmbH and Ko.KG).
For the alcian blue (AB) stain, sections were incubated for 3 min in 1% acetic acid. Then they were incubated for 30 min in 1% AB staining solution (Carl Roth GmbH and Ko.KG). After rinsing in 3% acetic acid and followed by a washing step in distilled water lasting 2 min, cell nuclei were counterstained with nuclear fast red aluminum sulphate solution (Carl Roth GmbH and Ko.KG) for 5 min. HE- and AB-stained sections were covered with Entellan (Merck KGaA). Images were taken using a light microscope (DM1000 LED, Leica Microsystems GmbH).
For hematoxylin-eosin (HE) staining, sections of sheets were incubated for 6 min in Harry’s hematoxylin (Carl Roth GmbH and Ko.KG), before being rinsed in running tap water and counterstained for 4 min in eosin (Carl Roth GmbH and Ko.KG).
For the alcian blue (AB) stain, sections were incubated for 3 min in 1% acetic acid. Then they were incubated for 30 min in 1% AB staining solution (Carl Roth GmbH and Ko.KG). After rinsing in 3% acetic acid and followed by a washing step in distilled water lasting 2 min, cell nuclei were counterstained with nuclear fast red aluminum sulphate solution (Carl Roth GmbH and Ko.KG) for 5 min. HE- and AB-stained sections were covered with Entellan (Merck KGaA). Images were taken using a light microscope (DM1000 LED, Leica Microsystems GmbH).
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