[Histonet] Poor quality frozen sections, feline skin.

[Peter Bodig]

I have some feline skin samples for frozen sections that were sent to us because our client was not able to section them themselves. Unfortunately we are having the same problems trying to make good sections ourselves. The samples seem dry and unsupported when we try to cut them. The tissue breaks into small pieces and falls out or the OCT when we try to cut it. We have tried fixation with 10% NBF for various times up to 24 hrs prior to doing frozen sections to give the tissue better support, which helps, but not enough, and then the tissue falls off the coated slides when we try to stain them. Sectioning at 8µ helps as well (that is as thick as out cryostat will go).

 

Does anyone have suggestions on how to get good quality frozen sections from feline skin?

 

We have come up with some questions as well:

 

1.	Is there a way to support the tissue by infiltrating it with OCT or other similar media to improve our sections?
2.	I have read about using 30% sucrose solution for cryo protection, does that aide in making good quality sections or just reduce freezing artifact?
3.	Does tissue need to be fixed prior to using cryo protection?

 

Our client will be using the sections for Senescence β-Galactosidase Staining.

 

Thank you for any help you can provide,

 

 

Péter Bodig

Laboratory Coordinator

Colorado Histo-Prep

E-mail: pbodig <@t> histoprep.com