Alcian Blue dye is used to prove glycosaminoglycan in mucins, for staining amyloids, cysteines and for polychromatic staining of mastocytes according to Alcian-Blue Safranin. It is also used to determine bacterial species and detecting bacterial capsules. Alcian Blue pH 2.5 kit enables adequate staining and visualisation of acid mucins without staining sulfated mucins. Rinsing solutions are of the same pH value, making the specificity of the reaction more intense. Cellular nuclei are stained red using a counterstain.
Alcian Blue pH 2.5 kit
Three-reagent kit for staining acid mucopolysaccharides according to Dorling and sulphated and carboxylated sialomucins blue. Slides are counterstained with Nuclear Fast Red (Kernechtrot) reagent to fully detect the presence of Alcian Blue positive staining.
Description
Additional information
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Stain Category | Carbohydrates |
PROCEDURE
Using kit for 100 tests | ||
1 | Deparaffinize the section in xylene (BioClear) or in a xylene substitute (BioClear New) | 3 exchanges, 2 minutes each |
2 | Rehydrate using 100% alcohol (Histanol 100) | 2 exchanges, 5 and 3 minutes |
3 | Rehydrate using 95% alcohol (Histanol 95) | 2 minutes |
4 | Rehydrate in distilled (demi) water | 2 minutes |
5 | Stain using Alcian Blue solution pH 2.5 (add ≥5 drops) | 45 minutes |
6 | Tilt the section and remove Alcian Blue solution pH 2.5. Without rinsing, cover the section with Sodium tetraborate, solution (add ≥5 drops) | 10 minutes |
7 | Rinse in distilled (demi) water | 5 minutes |
8 | Stain with Nuclear Fast Red (Kernechtrot) reagent (add ≥5 drops) | 5 minutes |
9 | Rinse in distilled (demi) water | 2 minutes |
10 | Dehydration using 70% alcohol (Histanol 70) | 5 dips |
11 | Dehydrate using 95% alcohol (Histanol 95) | 5 dips |
12 | Dehydrate using 100% alcohol (Histanol 100) | 2 minutes |
13 | Clear the section in xylene (BioClear) or in a xylene substitute (BioClear New) | 2 exchanges, 2 minutes each |
Immediately after clearing apply an appropriate BioMount medium for covering/mounting on the section. If BioClear xylene was used, use one of BioGnost's mounting xylene-based media (BioMount, BioMount High, BioMount M, BioMount DPX, BioMount C, or universal BioMount New). If BioClear New xylene substitute was used, the appropriate covering agent is BioMount New. Cover the section with a VitroGnost cover glass. |
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