Re: [Histonet] Iron leaching out during decal
1. Make certain that it is hematopoetic bone. That
is, bone with marrow (RBC, WBC, etc.). Vertebrae, rib, or sternum are fine.
But NOT adult femur heads, or bones from arms or legs from amputations from
adults. These are all fat and connective tissue. No iron stores in these bones
2. Use a mild decal for minimal time. We like to
use FASC (formic acid, sodium citrate) for 2-3 days. Stronger acids tend to
leach out the iron stores. If using hydrochloric or nitric acids, use 5% for a
few hours to overnight, depending upon the size (see #3).
3. Use minimum size. The larger the size you gross,
the longer it has to be in decal, and the more iron that gets leached out.
The HTL exam asks for bone 1 cm long. There is no requirement for width. So at
needle core biopsy would be acceptable, as long as there is cortex and
4. Realize that not all adults have large amounts
of iron stores. If you are getting it from an autopsy from someone who has been
sick a long time, or who has been undergoing treatment for cancer, they may have
very little to no iron stores. Try several different sources.
5. Make certain the formalin is OK. Acidic formalin
will also leech out the iron, so use newly made, buffered formalin. And fix for
minimal time. If the bone is being stored in formalin for a length of time
(weeks to months), the formalin can go acidic also.
Just some hints we've learned along the
Peggy A. Wenk, HTL(ASCP)SLS
School of Histologic Technicians and
William Beaumont Hospital
Royal Oak, MI
----- Original Message -----
Sent: Monday, August 18, 2003 3:27
Subject: [Histonet] Iron leaching out
Please help! I am working on my HTL Practical test and I am required to
do a Prussian Blue iron stain on a 1.0 cm piece of bone. My problem is that
everything that I use to decal my bone (so that it cuts nicely) leaches out
all of the iron. HCL decal solutions were obviously too powerful (tried them
anyways) so I used a milder selection that included acetic acid (after fixing
w/ formaldehyde), and a combination of formic acid and formaldehyde (to fix
and decal in the same step). I am leary to get Zenker's solution (which was
recommended by Frieda Carson's Histology: A Self Instructional Text)
because if it doesn't work, then I am left with the expensive and arduous task
of disposing of the mercury. Does Zenker's really fix and decal effectively to
preserve the iron AND allow me to cut a nice section? What about
Zinc Chloride to replace the mercury in Zenker's? Any suggestions would
be greatly appreciated, I really need some help. Thank you!
Bethany J. Krafels
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