[Histonet] Frozen sections missing centers

[Carla M Conway]

Hello,

I have been asked to to prepare frozen sections of fish livers that have 
been stored in RNAlater  Solution at -80C. Tissues (about 10 um square x 5 
um thick) were embedded in OCT, frozen on dry ice, and sectioned at 6 um. 
My problem is that when I cut a section, the center of the tissue is 
missing! Approximately 1 mm of tissue around the edges seems to section 
just fine, but then there is just a gaping hole in the center as if the 
tissue is receding while it is being cut. 

 I've tried sectioning at various temperatures (-20C, -16C, and -12C) and 
thickness (4um-8um). 

Any help will be greatly appreciated,


Carla Conway

Western Fisheries Research Center
Seattle, WA
ph: 206-526-6282 ext. 242
cmconway <@t> usgs.gov