Alcian Blue

Classification: Carbohydrate stain

Purpose: stain 
Group II & III Carbohydrates Sulfated and carboxylated acid mucopolysaccharides Sulfated and carboxylated sialomucins (glycoproteins)

Mechanism of staining: Ionic bonding or salt linkage

Control tissues: small intestine or large intestine

Alcian Blue (pH 2.5)

Mucopolysaccharides and sialomucins - dark blue 

Background - pink to red

Nuclear fast red counterstain 

Demonstrates both sulphated and carboxylated acid mucopolysaccharides and sulphated and carboxylated sialomucins at pH 2.5.

Small intestine


REAGENT

REAGENT

PURPOSE

MECHANISM OF STAINING

SOURCE OF ERROR

Alcian blue in 3% acetic acid

For pH 2.5 

Amphoteric dye

Demonstrate carboxylated & sulphated mucopolysaccharides and sialomucins

(progressively and directly)

Salt linkage - attaches to negatively charged acid mucopolysaccharides and sialomucins to produced an insoluble end product

 

Omitted: Omitted: target carbohydrates will be not be demonstrated.

Too short: Target will be lightly stained.

Too long: Background staining and darkly stained target tissue.

Alcian blue in water

For pH 1.0

Amphoteric dye

Demonstrate sulphated only mucopolysaccharides and sialomucins

(progressively and directly)

Salt linkage - attaches to negatively charged acid mucopolysaccharides and sialomucins to produced an insoluble end product

 

Omitted: Target carbohydrates will be not be demonstrated.

Too short: Target will be lightly stained.

Too long: Background staining and darkly stained target tissue.

Nuclear fast red

Counterstain

 

Omitted: Other tissue components not demonstrated.

Too long: Other tissue components stain darkly and obscure the primary stain.

Too short: Other tissue components stain lightly or are not demonstrated.

 

 

Alcian Blue (pH 1.0)

Periodic Acid Schiff and Hematoxylin counterstain   

Sulphated mucopolysaccharides - pale blue   

Background - pink to red 

Demonstrates sulphated mucopolysaccharides and sialomucins at pH 1.0


Alcian Blue 

Periodic Acid Schiff and Hematoxylin counterstain


Alcian Blue

Counterstain - Hematoxylin

 


Special Considerations

A variety of counterstains can be used including: Hematoxylin, Periodic Acid Schiff, Nuclear Fast Red, and Van Geison. 


Alcian blue is a basic and amphoteric dye. Adjusting the pH of Alcian blue alters the type of tissue it can bond with. At pH 2.5, both sulphated and carboxylated acid mucopolysaccharides and sialomucins are demonstrated. A
t pH 1.0, only sulphated mucopolysaccharides and sialomucins are demonstrated. 




References

Officer B. HIML251 Lecture notes: Alcian Blue and Masson Trichrome stains, January 28, 2009