Solarbio Alcian Blue Periodic Acid Schiff(AB-PAS) Stain Kit

Kit Components

Introduction

Glycogen staining is a conventional pathology staining method. First used by McManus in 1946, PAS technology displays mucin and other polysaccharides. This method is employed to visualize glycogen and other polysaccharides. The staining solution can also display neutral mucilaginous substances and some acidic substances.

The combination of Alcian Blue and PAS technology identifies neutral mucin and acid mucin in the same tissue section. This technique is commonly used to detect mucins. A negative staining result clearly indicates the absence of mucin. Typically, sections are first stained with standard Alcian Blue (pH 2.5) followed by PAS technology. Alcian Blue can dye salivary mucin, hiomucin, and proteoglycan blue. PAS technology can dye neutral mucin deep red or red-purple and simultaneously dye tissues and cells containing both neutral and acid mucin into different shades of purple, due to the combination and reaction of Alcian Blue and Schiff Reagent.

Self Provided Materials

Distilled water, Series of ethanol

Protocol (for reference only)

1. Dewax to distilled water, then rinse in distilled water for 2 minutes.
2. Stain with Alcian Blue Staining Solution for 10-20 minutes.
3. Rinse in distilled water three times for each time 1-2 minutes.
4. Treat with Oxidant for 5 minutes. Rinse in tap water and then rinse in distilled water twice.
5. Soak in Schiff Reagent and stain for 10-20 minutes. Then wash with running water for 10 minutes.
6. Stain with Hematoxylin Solution for 1-2 minutes, then wash with water.
7. Differentiate with Acidic Differentiation Solution for 2-5 seconds, then wash with water.
8. Blue with Scott Bluing Solution for 3 minutes and wash with water for 3 minutes.
9. Dehydrate with a series of ethanol, transparent with xylene, and seal with resinene.

Result

• Glycogen, neutral mucins, and various glycoproteins: Purplish Red
• Acidic mucins (sulfated and carboxylated): Blue
• Proteoglycans, hyaluronic acid: Blue

Note: Cells or tissues containing neutral and acidic mucins can be dyed different shades of blue-purple to purple.

Note

1. Ensure clean section dewaxing to avoid affecting the dyeing effect.
2. The oxidation time of Oxidant should not be too long; the optimal temperature is 18-22°C.
3. Reagents A, B, and C should be stored airtight at 4°C, avoiding excessive sunlight and air exposure. Before use, allow them to reach room temperature in the dark.
4. Acidic Differentiation Solution should be replaced frequently, and differentiation time adjusted based on section thickness, tissue type, and Solution age. Sufficient tap water washing after differentiation is essential.
5. The action time in Oxidant and Schiff Reagent is critical and depends on section thickness and tissue type.
6. When re-dyeing with Hematoxylin Solution, perform light dyeing to avoid masking alcian blue’s color. The goal is to prevent cytoplasm or mucin staining from covering alcian blue’s color.
7. The sequence of Alcian Blue-PAS Stain can influence final results. When PAS Staining precedes Alcian Blue Staining, neutral mucin and glycogen can be dyed purple. Conversely, Alcian Blue can dye these substances purplish red before PAS Staining.
8. For safety and health, wear experimental clothes and disposable gloves.