[Histonet] Remove OCT from frozen tissue block

[Benoît Delatour]

Dear histoneters,
We got mice brains that were initially sucrose-cryoprotected and then 
embedded in OCT - flash frozen and stored at -80°C. These tissues were 
initially prepared for subsequent cryostat sectioning but due to 
technical considerations we need to cut them using a sliding microtome 
to get thick (40µm) floating sections. We therefore would like to remove 
OCT before cutting. As OCT is water soluble it is expected that placing 
the OCT blocks in buffer would help removing the embedding media but 
thawing the tissue and then re-freezing it on the microtome stage might 
not be the best way to proceed (expected formation of ice crystals with 
known associated histological artefacts)...

We would appreciate any alternative solutions. Thanks!
Benoît


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Re: [Histonet] Remove OCT from frozen tissue block

[Paula Sicurello]

Hi Benoit,

You can just let the OCT thaw at room temperature then blot away the melted
OCT.  To speed up the thawing process carefully cut away as much of the OCT
as you can with a scalpel or razor blade and then let it thaw.

I hope this information is helpful.

Paula


On Thu, Oct 31, 2013 at 3:57 AM, Benoît Delatour benoit.delat...@upmc.frwrote:

 Dear histoneters,
 We got mice brains that were initially sucrose-cryoprotected and then
 embedded in OCT - flash frozen and stored at -80°C. These tissues were
 initially prepared for subsequent cryostat sectioning but due to technical
 considerations we need to cut them using a sliding microtome to get thick
 (40µm) floating sections. We therefore would like to remove OCT before
 cutting. As OCT is water soluble it is expected that placing the OCT blocks
 in buffer would help removing the embedding media but thawing the tissue
 and then re-freezing it on the microtome stage might not be the best way to
 proceed (expected formation of ice crystals with known associated
 histological artefacts)...
 We would appreciate any alternative solutions. Thanks!
 Benoît


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Paula Sicurello, HTL (ASCP)
Supervisor, Clinical Electron Microscopy Laboratory
Duke University Health System
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