more info again

From:becky riesen

 I have problem staining on my negative serum controls.  I purchase 
the negative solution from DAKO (prediluted) which works fine on 
everything except my Bone Marrows.  We used to fix Bone Marrows in 
Isotonic Buffered Formalin and Decalcify with a commercial decal 
from Surgipath (Decal I).  We had the problem then and we got a new 
pathologist who said where he was at they used Lerner D-calcifier 
and BBC's B-plus Fixative.  We've tried that and still have the problems.
The very specific Granulocytic WBC staining is found in both the 
bone biopsy and the aspirate, which makes me think the problem can't 
be in the decal.  On the same runs with the same negative mouse and 
negative rabbit solutions being used we have clean negatives on "tissues" 
and positive staining on the bone marrows, which makes me think it 
is something specifically about bone marrows.  Bone marrows are quite 
bloody, so I've increased the time to quench endogenous peroxidase 
(3%)from 6 minutes (which was routine for tissues) to 12 minutes.
Still have staining.  I know that if I get solutions too strong 
or go too long I'm going to cause other problems.  Hope the continued 
saga of info is helpful so that you all can help me.  

Naples Free-Net Web Mail

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