[Histonet] SAB for amyloid

[Tony Henwood]

The technique is as follows:

SULPHATED ALCIAN BLUE FOR AMYLOID


PRINCIPLE:

Amyloid has an increased avidity for alcian blue in the presence of added 
salt.  By fixing with an akali solution, SAB can be followed by various 
trichromic counterstains.

FIXATION:      10% buffered formalin

MICROTOMY:     7um 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.
     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.

5.   Van Gieson counterstain.

METHOD:

1.   Hydrate section 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 are also blue-green.

REFERENCE:

Lendrum, A.C., Slidders, W., Fraser, D.S., (1972).  "Renal Hyalin:  A Study 
of Amyloidosis and Diabetic Fibrinous Vasculosis with new Staining 
Methods".  J. Clin. Path. 25, 373-396.

This is from an old manual. Sorry that the Hazardous warning phrases are
missing.

Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at  Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318

http://www.histosearch.com/homepages/TonyHenwood/default.html
http://us.geocities.com/tonyhenwoodau/index.html




-----Original Message-----
From: rfail <@t> toolkitmail.com [mailto:rfail <@t> toolkitmail.com]
Sent: Friday, 19 September 2003 6:13
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] SAB for amyloid



Does anyone know of an amyloid stain with the abbreviation SAB?

Rena Fail 



_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


**********************************************************************
This email and any files transmitted with it are confidential and
intended solely for the use of the individual or entity to whom they
are addressed. If you are not the intended recipient, please 
delete it and notify the sender.

Views expressed in this message and any attachments are those 
of the individual sender, and are not necessarily the views of The
Children's Hospital at Westmead

This footnote also confirms that this email message has been 
virus scanned and although no computer viruses were detected,
the Childrens Hospital at Westmead accepts no liability for any 
consequential damage resulting from email containing computer 
viruses.
**********************************************************************