Hello, 
I am having trouble with sectioning of paraffin embedded human aortic valve 
samples. 
I received the embedded samples from a collaborator who had the embedding done 
by the pathology department at her institution. 

Six of the 12 samples sectioned fine but the remaining samples are developing 
ribbon cuts and/or are crumbling in areas when sectioned. The suggestion that 
calcification is causing the issue has been raised by other members of my lab. 
Some of the samples do appear to have calcific nodules while others may have a 
small bit of calcification along the valve. The samples with what appears to be 
calcific nodules do feel stiff at the nodules and are not being cut flat with 
the rest of the sample there. If I reposition the samples to a new part of the 
blade, the defects (ribbons or crumbling) are still in the same place or 
reappear in the same place quickly. 

I use Tissue-Tek Accu-Edge disposable high-profile microtome blades on a Lieca 
RM 2135 microtome.

For the samples that do not have the obvious nodules - is it possible this was 
an embedding error and I may be able to re-embed the samples and proceed?
If calcification is the issue - is it possible to de-calcify the valves after 
they have already been fix, dehydrated, and embedded? Or are there any other 
suggestions on how I could proceed?
Any advice or suggestions would be greatly appreciated. Thanks. 
Jen
--
Jennifer Petsche Connell, Ph.D.
Lab Manager and Research Scientist
Integrative Matrix Mechanics Laboratory <http://www.proteoglycan.com/>
Department of Bioengineering
Rice University


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