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<DIV><SPAN class=917260614-21102003><FONT face=Arial color=#0000ff
size=4>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.</FONT></SPAN></DIV>
<DIV><SPAN class=917260614-21102003><FONT face=Arial color=#0000ff size=4>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.</FONT></SPAN></DIV>
<DIV><SPAN class=917260614-21102003><FONT face=Arial color=#0000ff size=4>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.</FONT></SPAN></DIV>
<DIV><SPAN class=917260614-21102003><FONT face=Arial color=#0000ff size=4>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.</FONT></SPAN></DIV>
<DIV><SPAN class=917260614-21102003><FONT face=Arial color=#0000ff size=4>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.</FONT></SPAN></DIV>
<DIV><SPAN class=917260614-21102003><FONT face=Arial color=#0000ff size=4>The
staining suspension is made by adding 7 ml glacial acetic acid to 28 ml of the
dialyzed stock suspension.</FONT></SPAN></DIV>
<DIV><SPAN class=917260614-21102003><FONT face=Arial color=#0000ff
size=4>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.</FONT></SPAN></DIV>
<DIV><SPAN class=917260614-21102003><FONT face=Arial color=#0000ff
size=4></FONT></SPAN> </DIV>
<DIV><SPAN class=917260614-21102003><FONT face=Arial color=#0000ff
size=4>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.</FONT></SPAN></DIV>
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<DIV class=OutlookMessageHeader lang=en-us dir=ltr align=left><FONT
face=Tahoma size=2>-----Original Message-----<BR><B>From:</B> Gudrun Lang
[mailto:gudrun.lang@aon.at] <BR><B>Sent:</B> Tuesday, October 21, 2003 8:34
AM<BR><B>To:</B> Histonetliste<BR><B>Subject:</B> [Histonet] dialysed
ironsolution for Hale's<BR><BR></FONT></DIV>
<DIV><FONT face=Arial size=2>Dear histonetters!</FONT></DIV>
<DIV><FONT face=Arial size=2>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.</FONT></DIV>
<DIV><FONT face=Arial size=2>Please tell me the detailled procedure, when the
text say: "This mixture is dialysed against regularly changed water for three
days."</FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2>Gundi Lang</FONT></DIV>
<DIV><FONT face=Arial size=2>general hospital, Linz,
Austria</FONT></DIV></BLOCKQUOTE> <br> <br> <br>The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses.<br>Barry University - Miami Shores, FL (http://www.barry.edu) </BODY></HTML>