[Histonet] Perfusion fixation of pig lungs

LNLJ (Lene Lyngsie Jensen) lnlj <@t> novonordisk.com
Fri Jan 5 04:35:50 CST 2007

Hi Geoff, Kemlo, Edwards and histonet.

Thank you for your contribution. Unfortunately is fixation via the
trachea and bronchial tree not an option. We tried it in the initiating
studies on rats. Perfusion fixation via the vascular system worked in
rats (we did a full body fixation) but they are also significant smaller
than a pig.


It would be very good if we could get a fixation via the vascular system
in pigs as well, but limited to the lungs (because of the body size of
the pig). This fixation allows the airways and alveoli to remain I their
air filled state. At the same time allows this method fixation of the
surface lining layer of alveoli and smaller airways, which is washed
away in instillation fixation. 


The perfusion fixation is much more complicated, because it requires a
number of parameters to be controlled, such as the pressure applied to
the vessels, the degree of inflation of the air spaces, the
transpulmonary pressure, as well as the osmotic and oncotic pressure of
the fixative.


It would have been excellent if there was some one here that knew how to
do it in pigs.


I have also thought off (as Geoff also suggested) to limit the perfusion
to one lung at the time. To ensure a minimize variation I the fixation
from animal to animal.


The goal is also to find a fixation way that can work with PFA and
fixative used for EM as well. 


If any one can contribute with some more ideas, please do so.


Thanks to Geoff, Kemlo and Edwards.


Lene :-)



-----Original Message-----
From: Geoff McAuliffe [mailto:mcauliff <@t> umdnj.edu] 
Sent: 4. januar 2007 17:18
To: LNLJ (Lene Lyngsie Jensen)
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Perfusion fixation of pig lungs


Hi Lene:


1. Decide if you want to perfuse via the vascular system (washing out 

the blood) or instill fixative via the trachea and bronchial tree.  Note

that the position of the animal during instillation will determine which

regions of the lung are fixed first (or at all!). Study of the anatomy 

of the bronchial tree of the pig will pay dividends.

2. Decide if you are going to look at the entire lung or just one (or 

more) small regions. I suggest looking at one bronchopulmonary segment 

from one (or more?) lobes to conserve resources and minimize variation 

from animal to animal. If that is your decision then you can do a little

dissection and either perfuse that segment or instill into that segment.

    Good luck!




LNLJ (Lene Lyngsie Jensen) wrote:




>I hope that someone here can help me.


>We are planning to do some research in pig lungs. Due to this

>investigation it is very important to have a fast and effective



>We will therefore try to make a perfusion fixation. But a pig is a very

>large animal and therefore we would like to limit the fixation only to

>the lung region, and not a full body perfusion fixation.


>Is there anyone how has experience with perfusion fixation limit to

>one organ in pigs? 


>All suggestion will be read with enthusiasm.


>Experience from similar fixation methods in other animals is welcome as






>Lene Lyngsie Jensen


>Histonet mailing list

>Histonet <@t> lists.utsouthwestern.edu










Geoff McAuliffe, Ph.D.

Neuroscience and Cell Biology

Robert Wood Johnson Medical School

675 Hoes Lane, Piscataway, NJ 08854

voice: (732)-235-4583; fax: -4029 

mcauliff <@t> umdnj.edu




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