ALCIAN BLUE TECHNIQUE

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.
   

    

NOTE: No responsibility is assumed by The University of Nottingham or the Queens Medical Centre NHS Trust for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions or ideas contained in the material herein. It is the users responsibilty to ensure that all procedures are carried out according to appropriate Health and Safety requirements.