Prussian Blue rxn in bone, no postive rxn.


I am currently working on my HTL designation. One of the requested specimens 
for my practical is bone with the cortex and marrow. The bone is to be 
stained for iron with a Prussian blue reaction.

I have tried many different bone specimens using the Perls method with no 
positive reaction, but the control (liver) is positive. One specimen the 
pathologist believed he saw hemosiderin in the H&E, but with the Perls again 
being negative. :(
All of the glass ware was chemically clean. I used 10% Potassium Ferrocyanide 
and 10% HCL all fresh. I initially tried 5 minutes in Potassium Ferrocyanide 
and then 10 minutes with no results. I have used slow decal (formical) and 
overnight decal. Is the acid content in these products causing the loss of 
All of the specimens are animal (dog and min-pigs) . Would human bone exhibit 
more iron?
I would appreciate any suggestions to changes in processing or staining 

Loriann Hamilton
Powell, Ohio

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