I've been trying to get decent 10 micron sections of rat brains and the 
cutting is great but the brain tissue puckers when it hits the waterbath 
and I can't get a flat section on the slide.
The brain was perfused (not by me so this is a guess) with 
paraformaldehyde, stored in 0.1M phosphate/4% paraformaldehyde before it 
was brought to the lab.
I processed it on my little old rotary processor - one hour for each 
station beginning with 70% ETOH. I allowed extra time for infiltration 
since it was a thicker piece of tissue. I use paraplast xtra. This is the 
paraffin that I use routinely in the lab for all types of tissue.
Waterbath temp is 42 degrees. I use DI water and I even tried putting in a 
small amount of alcohol in the waterbath.
Any suggestions for flat sections? Should I raise the temp in the waterbath 
- I don't want the tissue to expand so fast that it "blows up".
At this point can the brains be re-infiltrated with a different type of 
paraffin? I'm thinking of a softer type.
Thanks!
Andi
.....................................................................
: Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
: Sr. Research Specialist       University of Arizona               :
: (office:  AHSC 4212)          P.O. Box 245044                     :
: (voice:  520-626-4415)        Tucson, AZ  85724-5044    USA       :
: (FAX:  520-626-2097)          (email:  algranth@u.arizona.edu)       :
:...................................................................:
           http://www.cba.arizona.edu/histology-lab.html