[Histonet] Iron stains

From:"Tom McNemar"

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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