[Histonet] collecting fresh tissue prior to perfusion

[Monfils, Paul]

Caroline,

I have done what you describe on a few occasions, not because I wanted to do
DNA analysis but because I wanted to do electron microscopy along with
standard histology. Cutting off a quarter of a lobe would probably allow
excessive leakage during subsequent perfusion, not only because of the
surface area involved, but also because you would undoubtedly cut some
fairly large vessels.  I don't know how much tissue volume you need for DNA
studies, but we needed only a tiny biopsy for EM (a couple of cubic
millimeters).  We cut a tiny notch from the edge of one lobe, using a small
scalpel, and the amount of capillary leakage from such a small peripheral
incision was negligible.

Paul Monfils

> ----------
> From: 	histonet-bounces <@t> lists.utsouthwestern.edu on behalf of
> Caroline Bass
> Sent: 	Wednesday, February 1, 2006 10:52 AM
> To: 	Histonet
> Subject: 	[Histonet] collecting fresh tissue prior to perfusion
> 
> Hello,
> 
> I have what may be a silly question but I thought maybe it is worth a  
> try.  I am in a situation where the best way to collect liver tissue  
> for my purposes is to perfuse the mouse with PFA.  However, on  
> occasion I need a small bit of fresh liver tissue to analyze for  
> DNA.  I was wondering if it is possible to collect a portion of a  
> liver lobe at some point in the perfusion process before I use  
> fixative?  For example, I could start the perfusion with heparinized  
> saline, clip a quarter of one lobe of the liver off and then switch  
> over to the PFA.  I would then proceed with the perfusion as normal.
> 
> The only complication I can think of is that the loss of a  
> significant portion of the liver could effect how well the rest of  
> the liver perfuses.
> 
> Has anyone tried this or is this something I should avoid?  I am  
> trying to cut down on the number of mice I am using.
> 
> My goal is to obtain a completely fixed liver lobe that I can section  
> all the way through.  It is my understanding that the mouse liver is  
> too large to fix by immersion in PFA, so it either has to be cut into  
> blocks or the mouse perfused.
> 
> Thanks,
> 
> Caroline
> 
>   
> 
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