[Histonet] paraffin sectioning problems

[Rene J Buesa]

It is likely that you have an infiltration problem and toluene twice for 15 minutes each is too little. Also your tissue subjects are regularly quite difficult. Go to any histotechniques manual and try to reproduce one of the methods in it. This would be a good starting point.
René J.

--- On Mon, 9/29/08, James Dooley <jdooley2008 <@t> yahoo.com> wrote:

From: James Dooley <jdooley2008 <@t> yahoo.com>
Subject: [Histonet] paraffin sectioning problems
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, September 29, 2008, 7:12 PM

I recently started paraffin sectioning and I am having trouble with getting good
quality sections reproducibly. I was wondering if you have any tips that may
help. I am sectioning thymus, pancreas, and lung. Section thickness is 10 um.
The microtome that I am using is Reichert-Jung Supercut, disposable knife holder
by Leica, and Accu-Edge low profile blades. I fix the tissue in a modified
Carnoy's fix overnight, dehydrate in graded alcohol, 100% ETOH, Toluene 2x15
minutes, Paraffin at 65 degrees overnight with 2X changes of paraffin. Embed and
store at RT. The paraffin I am using is Fisherbrand Paraplast X-tra Tissue
embedding medium. Any suggestion would be greatly appreciated. 

Thank you in advance,
James Dooley


      
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