ALCIAN BLUE TECHNIQUE

 

SECTIONS

4micron Paraffin wax sections
 

 

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SOLUTIONS

 

1. Alcian Blue pH 3.1
   • Alcian Blue 1g
   • 0.5% Acetic Acid 100ml

    

2. Alcian Blue pH 2.5
   • Alcian Blue 1g
   • 3% Acetic Acid 100ml

    

3. Alcian Blue pH 1.0
   • Alcian Blue 1g
   • 0.1M Hydrochloric Acid 100ml

    

4. Alcian Blue pH 0.2
   • Alcian Blue 1g
   • 10% Sulphuric Acid 100ml

    

5. 0.1% Nuclear fast red
   • Nuclear fast red 0.1g
   • Aluminium Sulphate 2.5g
   • Distilled water 100ml

    

 

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METHOD

 

1. Take sections to water
2. Stain in alcian blue solution pH 2.5 for 5 minutes
3. Wash well in water
4. Counterstain in 0.1% nuclear fast red for 5 minutes
5. Wash in water
6. Dehydrate, clear . Mount sections in DPX

RESULTS

• Mucins - Bright Blue
• Nuclei - Red

 

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NOTES

1. To further identify specific types of acid mucins you need to use the other pH solutions.
   

    

   • Strongly sulphated mucins - use pH 0.2 Alcian Blue
   • Weakly sulphated mucins - use pH 2.5 or pH 1.0 Alcian Blue

    

2. When using low pH solutions drain and blot dry instead of washing at stage (3) of the method.