[Histonet] zinc fixation form tibia

Tony Henwood AnthonyH <@t> chw.edu.au
Sun Sep 20 18:20:48 CDT 2009


Does the zinc-fix contain formalin?
If not then the tissue may not protected from the effects of
decalcification (even the mild EDTA decal you used)and so cells may be
leached/destroyed in the tissue.


Do you use enzyme retrieval? This will also affect the cells if formalin
fixation is not adequate. HIER may not be affected as much. 


Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of julia
Sent: Saturday, 19 September 2009 12:11 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] zinc fixation form tibia


Hello. Histonetters. 

I have recently used zinc fixed tibia and calvaria for the staining of
CD31. My CD31 does seem to have worked, however there seems to be marrow
loss and digestion of the marrow also. 


I zinc fixed the bones for 48 hours and then used EDTA pH 7 for
decalcification for 2 weeks. I am unsure whether I have fixed the bones
for too long, whether the decal has been affected by the zinc fix (as
the bones were "jelly-like" before processing) or whether the
decalcification needs shortening when using zinc fixation.


If anyone has any experience in using zinc fixation on tibia for Immuno
staining and can offer some advice that would be much appreciated.






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