[Histonet] dialysed ironsolution for Hale's

[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 [mailto:gudrun.lang <@t> aon.at] 
	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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Barry University - Miami Shores, FL (http://www.barry.edu) 
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