Re: [Histonet] Paraffin sectioning rat brain

From:Roger Moretz

Wow!  My procedure seems trivial, but here goes:

Ice the blocks by placing in wet ice (enough water to
wet the tissue) for about 10-15min; collect the ribbon
and float on a 46C water bath and pick up on either
Plus or uncoated slides (I prefer uncoated for brain).
 Airdry overnight, 60C oven for 10min (it's part of
the cycle on the Leica stainer) & stain with
hematoxylin & eosin.  I may occasionally get some
wrinkles but that is usually due to the use of the
coated slides.  One comment--leaving the brains on ice
too long can cause the tissue to expand too fast on
the waterbath.

Roger Moretz, Ph.D.
Dept of Toxicology
Boehringer Ingelheim Pharmaceuticals
Ridgefield, CT\

--- John Baker  wrote:

> Hello All,  Can someone give me any insight into the
> procedure for 
> getting good 7 micron paraffin sections?  I work
> with bone usually but 
> am doing work for a Neurosurgery group.  I process
> my slices of rat and 
> gerbil brain for 8 hours and then embed in Paraplast
> Plus.  I ice my 
> blocks before taking ribbons and then float the
> sections on an alcohol 
> 20% EtOH pre-float bath then transfer the sections
> to a 50 degree C 
> waterbath.  I use Superfrost Plus slides and heat
> them over night on a 
> 45 degree C warming table.  I do a Hematoxylin
> -Gills 3 and alcoholic 
> eosin stain.  The doctor is trying to detect small
> infarctions and 
> areas showing ischemic neuronal changes -
> eosinophilic cytoplasm and 
> nuclear changes such as pyknosis, karyolysis and
> karyorrhexis.  
> Cellular detail is good but I am getting fine
> wrinkles in the tissue.  
> Any suggestions in my method appreciated.  Thanks in
> advance.  John
> John A. Baker
> The University of Michigan
> Orthopaedic Research Laboratories
> Histology Unit
> 109 Zina Pitcher Place, 2218 BSRB
> Ann Arbor, MI 48109-2200
> 734-936-1635
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