RE: [Histonet] dialysed ironsolution for Hale's
Message
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.
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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