An arterial plaque was partially decalcified; one section was submitted for H&E stain.H&E stained slide showed the presence of calcium in the tissue; however, when stained with von Kossa; the result was negative. Then the very same slide was re-stained with H&E and once again the slide showed the presence of calcium. Why is it so?
Von Kossa stain works by reacting with the PO4 in CaPO4. Perhaps the type of calcium that you are seeing on H&E is a different type of calcium such as calcium oxalate. Von Kossa will only pick up the calcium phosphate.
I agree with Brandon; not every type of calcium stain with von Kossa; you can use another stain for calcium like Alizarin red stain; you find below a link to the features of this method:
www.ihcworld.com/_protocols/.../alizarin_red_s.htm
As far as I know, H&E does not stain calcium. The reaction you are assuming is related to calcium may be due to binding of haematoxylin to some protein associated with the site of calcium deposition - this would still be left in the tissue even after the calcium itself has been removed by demineralization.
try to stain a positive control slide for calcium, if it works, then you may not see calcium , it may be bacterial colonies. both of them stain blue with H&E so you may try Gram's stain.
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