HALE'S COLLOIDAL FERRIC OXIDE OR DIALYZED IRON PROCEDURE for acid
mucopolysaccharides.

SOLUTIONS:

29% Ferric Chloride Solution USP

Stock Muller's Colloidal Iron Oxide Solution - Bring 250.0 ml distilled
water to a boil.  While the water is still boiling, pour 4.4 ml 29% Ferric
Chloride solution and stir.  When the solution has turned dark red, remove
from heat and allow to cool.

Working Colloidal Iron Solution (prepare fresh)
Muller's colloidal iron (stock).....20.0 ml
Distilled water..........................15.0 ml
Glacial acetic acid.................... 5.0 ml
         Mix just before use.

12% Acetic Acid Solution
Glacial Acetic Acid...............12.0 ml
Distilled water.......................88.0 ml

5% Potassium Ferrocyanide Solution
Potassium ferrocyanide..........5.0 gm
Distilled water......................100.0 ml

5% Hydrochloric Acid Solution
Conc. hydrochloric acid...........5.0 ml
Distilled water........................95.0 ml

Potassium Ferrocyanide-Hydrochloric Acid Solution (prepare fresh)
Potassium ferrocyanide, 5%................50.0 ml
Hydrochloric acid, 5%.........................50.0 ml
          Mix just before use.

PROCEDURE
1. Deparaffinize and hydrate to distilled water.  Defat frozen sections and
smears.  Remove pigment as needed.
2. Rinse in 12% acetic acid solution, 3 min.
3.  Immerse in working colloidal iron solution, 1-2 hour.
4.  Rinse in 12% acetic acid solution, 4 changes, 3 min. each.
5.  Rinse 5 min. in running water and then in distilled water.
6.  Immerse in hydrochloric acid-potassium ferrocyanid solution, 20 min.
7.  Wash in tap water, 5 min.
8.  Rinse in distilled water.
9.  Counterstain in alum-hematoxylin (Harris or Mayer), 5 min: rinse and
differentiate as needed.     For red nuclei, use Kernechtrot (nuclear fast
red).
10.  Dehydrate, clear and mount.

Results
Acid mucopolysaccharides, sialomucins, connective tissue and their tumor
mucins, and acidic epithelial mucins (goblet cell mucins), carcinoma mucins
-- deep (Prussian) blue

Nuclei, cytoplasm -- Blue, red with Kernachtrot

Mast cell granule and some microbial walls -- blue

Hemosiderin -- iron-positive blue

Ref:  Linda Vacca,  Laboratory Manual of Histochemistry, 1985

Hope this helps!!
Michelle Skelton
Alaska Native Medical Center

> -----Original Message-----
> From:	Frank Hogg [SMTP:FHogg@dchsystem.com]
> Sent:	Wednesday, September 08, 1999 7:08 AM
> To:	'histonet@pathology.swmed.edu'
> Subject:	Hale's Colloidal Iron Stain
> 
> Does anyone in histoland have a procedure for Hale's Colloidal Iron
> reaction. We have looked in both old and new AFIP manuals,
> Sheehan-Hrapchak,
> both Carson manuals and no Hale's can be found. Can someone help us out?
> Thanks.