Dear Sharon,
try putting your block on a slab of ice for 15mins instead (You can even
incorporate up to 4% Potassium or Sodium Hydroxide in your ice block to
further soften). As for the patchy  nuclei, autolysis is probably to blame,
but as a last resort use the Celstine/ Mayers Haemalum sequence or fresh
Weigert's. Of these the Celestine gives the better and sharper results, and
if this doesn't work, then you've got Buckley's and Nunn (Strine for no
hope).
Good luck, Mike Rentsch (Downunder)
-----Original Message-----
From: Marshall, Sharon, <marshall@anat.uct.ac.za>
To: histonet@pathology.swmed.edu <histonet@pathology.swmed.edu>
Date: Thursday, 9 March 2000 11:19
Subject: tendon/bone


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