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From:"Marshall, Sharon," <marshall@anat.uct.ac.za>
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Dear histonetters,

I need some advice on the cutting and staining of tendon/bone 
junctions.   I am busy sectioning archille's tendon and it's 
associated bone (calcaneus)  The specimen was taken from a human .
I am not sure of the post mortem interval.  The specimen was removed 
from the body and fixed in 10% neutral buffered formalin. After 
adequate fixation it was then decalcified in a HCL/E.D.T.A. mixture
,dehydrated, cleared and embedded in wax. I find the tendon quite 
tough to section. I have found that a cold water soak helps. Are 
there any other tricks to cutting this type of tissue? 
I have also found that on staining with Haematoxylin and Eosin that 
the tendon and  it's associated fibrocartilage are not staining up very 
well.  The tendon stains in a patchy way and the fibrocartilaginous 
zone is virtually unstained. Help!   The bony  areas are taking up 
the eosin quite well  although the nuclei of the cells within the 
bony areas are more eosinophilic then basophilic.  Could these 
problems be due to post mortem interval, could it be due to the 
decalcification or could it be the choice of Haematoxylin. I am using 
Mayer's.  Should I perhaps be using Harris's? I normally have no 
problems with other H&E's on other types of tissue. 
Sharon Marshall
Dept. of Anatomy & Cell Biology
University of Cape Town
E-mail: marshall@anat.uct.ac.za

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