Re: Hale's Colloidal Iron Stain

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From:Victoria Baker <>
To:Frank Hogg <>
Date:Wed, 08 Sep 1999 17:14:31 -0700 (PDT)
Content-Type:text/plain; charset=us-ascii

Hi Frank,

About a year ago I drove poor Russ Allison nuts with this request, only
to find totally by accident the procedure in Culling, the 3rd edition. 
The procedure really isn't as complex as it sounds.  

page 278

Hale's Collodial Iron Technique

"Hale's technique is based upon the affinity of acid groups for
colloidal iron at a low pH.  The iron forms a chelate with the acid
groups and may then be demonstrated by the Prussian Blue reaction.
While this method is excellent for the demonstration of acid mucins it
is inferior to Alcian blue as a specific method for histochemical

Special Solutions required

  Dialysed iron (British Drug Houses or Merck's)  1 volume
2M acetic acid       1 volume

Dialysed IronSolution (Rinehart & Abu'l Haj)
Dissolve gradually 75 g ferric chloride in 250 ml distilled water
adding 100 ml glycerol, followed by the gradual addition of 55 ml of 28
per cent ammonia with constant stirring.  This mixtured is dialysed
against regularly changed distilled water for 3 days.

Acid-ferrocyanide Solution
Equal parts of 2 per cent potassium ferrocyanide and 2 per cent
hydrochloric acid.

1.  Bring sections to water.
2.  Flood with dialysed iron solution (either solution works well) for
10 minutes.  ((Here I think he is referring to either the purchased
solution or home-made stock))
3.  Wash well with distilled water.
4.  Flood with acid-ferrocyanide solution for 10 minutes.
5.  Wash well in water.
6.  Counterstain lightly with 0.1 per cent neutral red or safranin.  
7.  Rinse in water and dehydrate rapidly.
8.  Clear in xylol and mount in DPX or HSR resin.


Acid mucopolysaccharides...........bright blue
Other tissue constituents..............shades of red"

I have copied this completely from the text, and I hope it helps you. 
Good luck.

Oh and Russ if you see this, I want you to know, that I appreciated
your help so much when I didn't have a clue what the pathologist was
looking for.  Finding this was a LOT too late for me.

Vikki Baker

--- Frank Hogg  wrote:
> 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.

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