[Histonet] damage to freeze-sections after cutting

[Gerben Schaaf]

Dear histonet users,

I am just starting to do cryosections and I ran into a problem. I have mouse tissue that we freeze in OCT (we tried different tissue preprocessing: PFA immersion fix, whole animal PFA perfusion, sucrose gradient). I am sectioning skeletal muscle at 10 micron. 
When I look at the sections directly after sectioning (slide is still cold), the morphology looks ok. However, if I allow the slide to warm to room temp than the tissue start to 'tear' apart and the individual fibers get separated. I could keep the sections at -20C and they will be ok, however, during staining the slides will warm to RT and the above problem occurs.

What can I do to prevent the tissue from starting to tear?
 

Thanks, 

Gerben, Rotterdam, The Netherlands


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