[Histonet] Finding iron in decalcified sections

Rene J Buesa rjbuesa <@t> yahoo.com
Thu Apr 23 08:20:11 CDT 2009

The mandible and maxilla of chicken's' beaks are porous bone covered by horny keratin so it is very likely that you will have to do a double procedure.
First to soften the covering keratin with 10% sodium or potasium hydroxide aq. solution and later decalcify the bone.
Since it is a poroues bone  you will be able to decalcify it with EDTA, as is done with bone marrow core biopsies, and if you do it that way, the iron will not be removed, as it is not removed from the core BM biopsies.
René J.

--- On Wed, 4/22/09, Farish, Craig <cfarish <@t> csu.edu.au> wrote:

From: Farish, Craig <cfarish <@t> csu.edu.au>
Subject: [Histonet] Finding iron in decalcified sections
To: histonet <@t> lists.utsouthwestern.edu
Date: Wednesday, April 22, 2009, 7:14 PM

Hi folks - I'd very much appreciate some help with this problem. I've
been asked to performing the routine histo for a trial investigating the
ability of chickens to sense direction. They are believed to have areas
of iron rich tissue in their beaks which can detect the magnetic field
of the earth in the same fashion as homing pigeons. The problem is this
- to section the beaks I need to decalcify them, however I'm pretty sure
that standard decalcification removes iron, at least partly. I know that
the levels of iron in the tissue are very low and I've just trialled my
technique on a pigeon with no success. I'm using a standard Perl stain
(2% HCl + 2% K Ferrocyanide for 20 mins) on FFPE tissue which usually
works very well for me, fixation is for a minimum of 24 hours in either
10%NBF or Bouins and decalcification is in either 5% Nitric acid or
formic acid. Would anyone have any suggestions as to how to decal
without removing iron or another stain which I could use in this

Thanks in advance for any and all suggestions,



Craig (Joe) Farish

Senior Technical Officer

Veterinary Diagnostics

School of Animal and Veterinary Sciences

Charles Sturt University

Wagga Wagga, Australia

c farish <@t> csu.edu.au


' I don't want to achieve immortality through my work, I want to
immortality through not dying' - Woody Allen


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