[Histonet] AB-PAS for Barrett's Esophagus

[Jason McGough]

1% Periodic Acid Solution                                       0.5% Sodium
Metabisulfite
 Periodic Acid....1 gm                                         Sodium
metabisulfite.0.5 gm
 Distilled water..100 ml                                         Distilled
water....100 ml

.1N HCL                                                                   3%
Glacial Acetic Acid
Conc. (10N) HCL.1 ml                                         Conc. Glacial
Acetic Acid..3 ml
Distilled Water.100 ml                                          Distilled
water......100 ml

Alcian Blue Solution  pH 1.0                                    Alcian Blue
Solution pH 2.5
Alcian Blue 8GX...1.0 gm                                     Alcian Blue
8GX....1.0 gm
0.1N HCL....100 ml                                      3% Glacial Acetic
Acid.100 ml
                                                                            
      Adjust pH to 2.5.  Add a few
Schiff's Reagent
Thymol crystals for preservative.
Purchased Commercially (ready to use)

PROCEDURAL NOTES
1.	pH 2.5 is routinely used in our laboratory. Use pH 1.0 only if the
pathologist specifically requests you to do so.
2.	Filter Alcian Blue solutions before use.
3.	Use an aliquot from the stock bottle of Schiff's reagent.  This may be
reused but do not return to stock bottle.  Change frequently and discard if
solution is pink.

PROCEDURE
1.	Deparaffinize and hydrate to distilled water.
2.	Place slides in freshly filtered Alcian Blue solution for 30 minutes.
3.	If using Alcian Blue solution 2.5 - Wash in running water for 5 minutes.
	If using Alcian Blue solution 1.0 - Blot section dry with fine filter
paper.
4.	Oxidize in Periodic Acid solution for 10 minutes.
5.	Wash in running water for 5 minutes.
6.	Place slides in Schiff's reagent for 10 minutes.
7.	Rinse in Sodium Metabisulfite solution, 3 changes; 2 minutes each.
8.	Wash in running water for 10 minutes.
9.	Dehydrate in 95% alcohol, 100% alcohol, clear in xylene, two changes
each.
10.	Coverslip.



RESULTS
1.	pH 2.5 (acid group)
-Exclusively acid substances (various connective tissue mucins) - blue
-Neutral polysaccharides (glycogen) - magenta
-Both Alcian blue and PAS, yielding varying shades of purple to deep blue,
color most epithelial mucins and cartilage ground substance.
-Cell bodies of fungi - red to purple
-Mucoid capsules - blue
-Other features resemble PAS stain
2.	pH 1.0 (sulphate group)
Sulfated mucosubstances - blue


REFERENCE
1.	Lev, R., Spicer, S.S.;J. Histochem. Cystochem. 12:309,1964.  Copyright by
Williams and Wilkins Co.
2.	C.F.A. Culling; Handbook of Histopathological and Histochemical
Techniques, 3rh e. 1974.  Butterworth.
3.	Preece, Ann: Manual for Histologic Technicians 2nd ed. 1965.
4.	AFIP Manual For Histologic Stain Methods, 3rd ed.
5.	Schenk, E.A., M.D. and Mowry, R.W., MD>D, Journal of Histotechnology Vol.
6#2, June 1983.


Jason McGough HT(ASCP)
Account Representative - Anatomic Pathology
Clinical Laboratory of the Black Hills
2805 5th Street Suite 210
Rapid City, SD 57701
605-343-2267 Ext 127
605-718-3779 (Fax)
jmcgough <@t> clinlab.com



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Diana
McCaig
Sent: Thursday, June 24, 2010 11:48 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] AB-PAS for Barrett's Esophagus


Can anyone share with me their procedure for AB-PAS for Barrett's
Esophagus.
With thanks

Diana
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