[Histonet] Undecalcified sample in paraffin and plastic media

[Rui TAHARA]

I have undecalcified paraffin embed samples
that were sectioned at 10 micron that I want to stain with Von kossa. Because
samples are embryonic quail heads (ossification starts to happen) and still
soft enough to section with standard rotary microtome with tungsten knife in paraffin.


My intention is to 3D reconstruct anatomies
based on histological sections. Because of this, I am wondering if I should actually
use plastic media rather than paraffin to keep the section shape as consistent
as possible. Does plastic embed material actually preserve the consistent shape
among sections better than paraffin embed sample? No winkle etc..? Is there any
other advantage that I actually should use the plastic media than paraffin for what
I want to do? I know downside of plastic media is that in general plastic
embedding process are lengthy and plastic embedding material are expensive than
the paraffin ones, and are mainly use for bone to support the hard material for
sectioning. 

When I sectioned some ossified samples, beak
start to fall off from section and the section show the lines from the possibly
scratched knife. Is this indication of paraffin media that does not provide enough
strength for sectioning? I thought it may possibly the poor infiltration. 

 

In our lab nobody has processed the plastic
embedding and sectioning (we have only standard microtome, no vaccum machine. Can
I section plastic embed sample with the standard microtome at 10 micron?) so I would
like to have any input before actually making a plastic embed sample. Any
suggestions would be appreciated. 


Rui TAHARA
Biology Department 
McGill University