Alcian Blue pH 2.5 kit

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)

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)

7

Rinse in distilled (demi) water

8

Stain with Nuclear Fast Red (Kernechtrot) reagent (add ≥5 drops)

9

Rinse in distilled (demi) water

10

Dehydration using 70% alcohol (Histanol 70)

5 dips

11

Dehydrate using 95% alcohol (Histanol 95)

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.