RE: [Histonet] Iron stains

From:"Weems, Joyce"

I would always fix first before decal. The acid is stripping the iron.
After discontinuing B5 fixative, we use B-Plus (zinc), but some are very
happy with 10% NBF. The problem with purchased controls is that they are
not treated the same as your material, but at least you know your
reagents are good!

My 2 cents... J:>)

Joyce Weems
Pathology Manager
Saint Joseph's Hospital 
5665 Peachtree Dunwoody Rd NE
Atlanta, GA 30342
Please note new phone and fax numbers
678-843-7376 - Phone
678-843-7831 - Fax

-----Original Message-----
[] On Behalf Of Tom
Sent: Monday, November 24, 2008 12:07 PM
Subject: [Histonet] Iron stains

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

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

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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