Sample Type Required: Deparaffinized and re-hydrated tissue sections on positively charged slides.
Fixative: 10% Neutral Buffered Formalin or Bouin Solution
Alcian Blue, pH 2.5
| Step | Reagent | Time | Technical Notes |
| 1 | 3% acetic acid (15mL glacial acetic acid and 485mL distilled water) | Three minutes | This rinse will protect the subsequent Alcian Blue Solution from pH changes due to the introduction of water. |
| 2 | 1% Alcian Blue Solution, pH 2.5 (5g Alcian Blue-8GX dissolved in 500mL 3% Acetic Acid) | 30 minutes at room temperature or 15 minutes at 37°C |
|
| 3 | 3% acetic acid (15mL glacial acetic acid and 485mL distilled water) | Brief rinse | Rinse well enough to remove excess Alcian Blue. This will help to prevent nonspecific staining. |
| 4 | Running tap water | Ten minutes |
|
| 5 | Distilled water | Rinse well |
|
| 6 | Nuclear Fast Red Solution | Five Minutes |
|
Expected Results
- Weakly acidic sulfated mucins = Dark blue
- Hyaluronic acid = Dark blue
- Sialomucins = Dark blue
- Background = Pink to red
- Nuclei = Red
Post Staining Procedure: Tissue sections should be rinsed well in distilled water, dehydrated with 95% and absolute alcohols, cleared and cover-slipped.
