[Histonet] rescue sections for staining

From:"J. Douglas Swarts"

We are trying to stain some sections cut about ten years ago.  They are 
rat cranial bases including the middle ear which is the area we are 
interested in.  When we stain the sections with H&E we get no 
hematoxylin staining, ie. no nuclei at all, but the eosin gives a nice 
uniform pink color. 

The specimens were decalcified in 10% formic acid which was neutralized 
with sodium sulfate according to the histologists notes.  The sections 
stained at the time show very little hematoxylin staining.  So the 
questions are:

Are these sections overdecalcified?  If so is there any technique I can 
use to rescue them?

Did the neutralization with sodium sulfate create the problem?  If so 
can I do anything to reverse the effect?

Finally, if one or the other of these situations are true and cannot be 
remedied is there another staining protocol that will give me a 
"reasonable" level of morphologic detail?

Thanks for your help.


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