Re: Lung Embedding Protocol

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From:Katie B <> (by way of histonet)
To:histonet <>
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---sbledsoe <> wrote:
> I have been asked to Post the following:
> Could someone please suggest a good protocol for embedding lung
tissue in
> paraffin.
> Thanks
> Sharon B. Bledsoe
> Indiana University
> School of Medicine
> Dept. of Surgery
> Indianapolis, IN 46202
> sbledsoe@iupui.ed

We routinely work with whole lungs from rat and mouse (and sometimes
the occasional horse lung lobe).  The key to getting good sections is
proper fixation which involves inflating the lung with fixative
through the trachea so that the avolei can be microscopically
examined.  The proper way to do this is by constant pressure perfusion
for about an hour for formalin fixation.  The most basic version of
the gizmos we have designed to accomplish this is a seperatory funnel,
filled with fixative, with tubing attached at the base so that a
cannulated lung can be attached.  The volume in the seperatory funnel
is kept constant so that the pressure in the lung will be constant.
Our protocol calls for 20-30 cm of pressure for rat and mouse lungs.
After fixation we then trim out the desired tissue region then submit
for routine paraffin embedding.

Catherine "Katie" Bresee Bennett
Laboratory for Experimental Pathology
Department of Veterinary Pathology
Michigan State University

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