Hi,
    I believe your eosinophillic nuclei are caused by the decalcificaion.
Have you tried formic acid? HCl is a little harsh, even with the EDTA if
the tissue is taking a long time to decalcify. The patchy staining may also
be an effect of this but it could also be from slight variations in
thickness of the section, or the tissue may need to be cleared and
infiltrated a little more in processing.
good luck, this sounds like a bear of a project
Amos Brooks

"Marshall, Sharon," wrote:

> 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.
> Thanks
> Sharon Marshall
> Dept. of Anatomy & Cell Biology
> University of Cape Town
> E-mail: marshall@anat.uct.ac.za