RE: [Histonet] dialysed ironsolution for Hale's

From:"Smith, Allen"

Dialysis is the movement of solutes through a semi-permeable membrane.  The objective is to eliminate particles too small to be useful in the staining reaction.
To make dialyzed iron one pours the ferric hydroxide suspension into seamless cellulose tubing with 2.4 nm pore radius (i.e. 4.8 nm pore diameter) and ties off the end of the tubing. (The other end was tied off before you began to pour.)  One immerses the tubing in distilled water for 3 days, changing the water 9 times a day.
To make 532 ml of colloidal iron suspension you will need about a 55 cm length of 23.5 mm radius (i.e., 47 mm diameter) seamless cellulose tubing, which you can tie off 5 cm from each end.  You should dialyze against about 15 litres of distilled water.
After the dialysis step one filters the suspension through Whatman #1 (medium porosity filter paper) to eliminate particles that are too large to be useful.
The ferric hydroxide suspension is made by dissolving 100 g ferric chloride hexahydrate in 333 ml distilled water and adding 133 ml glycerol.  One then adds 4 aliquots of concentrated ammonium hydroxide: 33 ml, 17 ml, 10 ml, and 6.2 ml; stirring until the precipitate is resuspended after each addition.
The staining suspension is made by adding 7 ml glacial acetic acid to 28 ml of the dialyzed stock suspension.
REFERENCE: J.F. Rinehart & S.K. Abul-Haj (1951) An improved method for histologic demonstration of acidic mucopolysaccharides in tissues. Arch. Pathol. 52: 189-194.
Personally, I think that dialyzing the ferric hydroxide is a lot of work for little advantage.  I usually use Mowry's quick and dirty method which can be found in Humason's "Animal Tissue Techniques."  The quick and dirty method works well 90% of the time.  ORIGINAL REFERENCE: R.W. Mowry (1958) J.Clin.Invest. 7: 566-576.
-----Original Message-----
From: Gudrun Lang []
Sent: Tuesday, October 21, 2003 8:34 AM
To: Histonetliste
Subject: [Histonet] dialysed ironsolution for Hale's

Dear histonetters!
I need some advice with the Hale's stain. I have found the procedures in the archives, but unfortunately I have no idea what "dialysed" in German means.
Please tell me the detailled procedure, when the text say: "This mixture is dialysed against regularly changed water for three days."
Gundi Lang
general hospital, Linz, Austria

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