[Histonet] Bad sections

[Rene J Buesa]

Compressed sections usually are the result of:
1- blunt knife
2- too soft paraffin for the selected thickness
3- too warm paraffin (block not cold enough)
4- blade too vertical (not enough clearing angle)
5- cutting too fast, specially with thin sections
6- too warm blade
7- microtome set screws loose or defective.

René J.


--- On Wed, 3/11/09, Va Paula Sicurello <vapatpxs <@t> yahoo.com> wrote:

From: Va Paula Sicurello <vapatpxs <@t> yahoo.com>
Subject: [Histonet] Bad sections
To: "HistoNet" <histonet <@t> lists.utsouthwestern.edu>
Date: Wednesday, March 11, 2009, 2:29 PM

Hi Histo netters,

I am am my wit's end.  I know how to section but my sections are
compressing like crazy.  I've altered my processing protocol thinking that I
was overprocessing my samples.

I've tried different knife angles, different brands of razor blade knives.

It's either me or the microtome.

Info- I process animal tissues with between 30-60 minute steps.  Vacuum
paraffins for 30 minutes.  Section with Reichert-Jung 2030 set at 4-ish degrees
(a little below the middle line on the knife holder).
Soak blocks on Downey ice blocks, water bath at 44 degrees.

Suggestions?  

Waa,

Paula

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


      


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