[Histonet] Sulfated Alcian Blue

[Tony Henwood]

This is our technique:




Sulphated Alcian Blue for Amyloid


 

Amyloid has an increased avidity for Alcian blue in the presence of added salt.  By fixing with an alkaline solution, SAB can be followed by various trichromic counterstains (Lendrum et al 1972 J Clin Pathol 25: 373-396).

 

Fixation:      10% buffered formalin

 

Microtomy:     7μm paraffin sections

 


Solutions:


 

1.      Acetic/Alcohol Rinse:

 

               95% ethanol                          45ml

               Distilled water                        45ml

               Glacial Acetic Acid                10ml

 

2.      Stock Solutions:

 

a)                  1% Alcian Blue (CI 74240) in 95% ethanol.


Caution Flammable - see MSDS


 

b)                  1% Sodium Sulphate Hydrate in distilled water.

 

3.      SAB Working Solution - prepare fresh:

 

               Stock solution (a)                  18ml

               Stock solution (b)                  18ml

               Glacial Acetic Acid                4ml

 

               Mix, stand 30 minutes before use.

 

4.      80% ethanol saturated with borax.


Caution Flammable - see MSDS


 

5.      Van Gieson counterstain.

 

 


Staining Procedure:


 

1.      Dewax, hydrate sections and wash for 10 minutes.

 

2.      Rinse in acetic/alcohol, 2 minutes.

 

3.      Place in SAB, 2 hours.

 

4.      Rinse in acetic/alcohol, 2 minutes.

 

5.      Wash in water.

 

6.      Alkalinise in borax/alcohol, 30 minutes.

 

7.      Wash in water.

 

8.      Counterstain in Van Giesons.

 

9.      Blot dry, dehydrate, clear and mount.

 

 


Results:


 

Amyloid

Blue-green

Collagen

Red

Muscle, cytoplasm

Yellow

Mast cell granules and some colloids 

Blue-green

 



Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Nathanial nauss
Sent: Saturday, 28 February 2009 5:42 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Sulfated Alcian Blue


I need some help, has any one used the sulfated alcian blue to stain for amyloid.  We have a case that looks like it should be positive but it is not staining with the Congo Red.  Any help would be great. Nathaniel _______________________________________________
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