After dewaxing and re-hydration, 5 µm thick sections were stained with hematoxylin/eosin (HE) to evaluate the morphological features. Other sections were stained with Periodic acid–Schiff (PAS) or with Alcian Blue pH 2.5 (AB) for histochemical purposes. Subsequently, new slides, were stained with Periodic acid–Schiff (PAS)/Alcian Blue pH 2.5 staining kit (Bio Optica, 04-163802) to analyze the overall complex carbohydrates [11 (link),12 (link)].
Lendrum’s staining was used to identify acidophilic granules containing cells. Sections were incubated with Phloxine B to visualize acidophilic compounds and then with Tartrazine to remove the non-specific staining.
The alkaline phosphatase expression was used as a marker for the identification of fully differentiated epithelium. Briefly, slides were rehydrated and brought to distilled water, were then immersed in fresh Tris HCL (pH 9.5) solution to create the alkaline environment for 5 min. They were then incubated with BCIP/NBT substrate (5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium, (Vector Laboratories, SK-4500 USA), which produces an indigo reaction product in the presence of alkaline phosphatase (AP) enzyme. Sections were then rinsed in tap water, counterstained using Mayer’s hematoxylin, dehydrated and mounted.
Lendrum’s staining was used to identify acidophilic granules containing cells. Sections were incubated with Phloxine B to visualize acidophilic compounds and then with Tartrazine to remove the non-specific staining.
The alkaline phosphatase expression was used as a marker for the identification of fully differentiated epithelium. Briefly, slides were rehydrated and brought to distilled water, were then immersed in fresh Tris HCL (pH 9.5) solution to create the alkaline environment for 5 min. They were then incubated with BCIP/NBT substrate (5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium, (Vector Laboratories, SK-4500 USA), which produces an indigo reaction product in the presence of alkaline phosphatase (AP) enzyme. Sections were then rinsed in tap water, counterstained using Mayer’s hematoxylin, dehydrated and mounted.
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