OH NO!  WRINKELED BRAINS!!!???  i thought all brains had those funny wrinkles
*vbg*  just kidding.

Seriously...

I section a lot of brain from all kinds of animals.  In my waterbath I put
approx 100 mL 95% ETOH in approx 1800 mL DI water.  I keep the water temp
around 35-ish (cool).  i pick up my section, place it on a slide warming tray--
set at about 45-50 C.  Before placing the slides on the warming tray, I slice
off any excess paraffin that overhangs the sides of the slide with a razor, or
a disecting probe.  I like the waterbath to be coolish because it makes it
easier to break apart the ribbon to get to a good section and I can manipulate
the section easier.  I have also found that the sections don't fall off during
staining if I mount  them to the slide in this way.   I have found the thing
that really beats those pesky wrinkles is free movement of the section (that's
why I remove the overhanging paraffin) and heat.  hope this helps.

Connie McManus

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               :
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> : (voice:  520-626-4415)        Tucson, AZ  85724-5044    USA       :
> : (FAX:  520-626-2097)          (email:  algranth@u.arizona.edu)       :
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>            http://www.cba.arizona.edu/histology-lab.html