I also would be interested in the responses to this dilemma. We have pulled 
out our hair on several occasions. I am thinking it is the paraformaldehyde 
or in my case, gluteraldehyde. This seems to be a regular occurrence for 
us  when we try to section rat brains.




At 09:25 AM 10/19/2000 -0700, Andrea Grantham wrote:
>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
>


**************************************************************************
Jill Songer HT (ASCP)
Anatomic Pathology Lab Supervisor
Veterinary Teaching Hospital
Virginia Tech
Blacksburg, Virginia 24061