[Histonet] Iron stains

From:"Amos Brooks"

   I'm betting your control and test slides are right, but there is a fairly
quick & dirty method of making sure. Don't process ALL the aspirates with
the rest of the case. Make some smears of them directly. Do try to pick up
some of the smaller spicule material (centrifuge if necessary) and when you
place it on the slides crush them up with edge of the smearing slide. Make
the smears and let them air dry (don't worry, you're looking for iron here
not necessarily morphology). Once they are dried place them gently in
methanol (I'm not really sure why we did this since it was air dried
already) and re hydrate to distilled H2O. Then run your usual Iron stain
with a control of course. This should confirm the results of the processed
material. If this is positive and your processed material is not THEN you
can panic. This should at least alleviate the Dr's concerns. If/when you
find a nice positive one (hemochromatosis ?sp?) take about 20-50 slides and
sit down and make a bunch of smears to store in a slide box for future
controls (the Iron isn't going anywhere).

Best of luck,

Message: 12
Date: Mon, 24 Nov 2008 12:06:59 -0500
From: "Tom McNemar" 
Subject: [Histonet] Iron stains
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We may be having a problem with our iron stains.  I say that we 'may' be
having a problem because our positive control always works.

For bone marrows, we stain the core, aspirate, a smear, and a purchased
positive control.  The stain is all over the place.  We may have a loaded
smear with a negative core and aspirate.  We may have a positive aspirate
with a negative core and smear.  We may have all negatives even though
clinically, there should be iron.  Our control always works.  The
pathologists do not trust the stain and really do not believe the results.
 I have stood by the stain due to the positive control but I find it
increasingly difficult.

I would appreciate any thoughts.  Our procedures are outlined below...

       The aspirate is allowed to clot before placing it into 10%NBF.
       The core is placed briefly into DecalStat (until it floats) then
placed into 10%NBF.
       Both are then processed with our regular tissues.  The spend anywhere
from 4 to 10 hours in formalin.
       The following day, they are cut and along with one of the purchased
controls, run down to water on the stainer.  (8 mins onboard
 oven, americlear, alcohols, water)
       The smear is placed into 95% alcohol then rinsed in distilled.
       All slides are then stained using Perl's Method for iron pigment.

We have already explored the way the specimens are collected during the BM
procedure, and decal solution.  All reagents are good.  I'm wondering about
the formalin fixation.  I know that threre are other probably better
fixatives.  Does anybody use another fixative prior to formalin?  For those
using formalin only, do you limit the fixation time in any way?

I really would appreciate any comments.  Such a common and simple stain.  We
should be able to trust it.


Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
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