RE: [Histonet] collecting fresh tissue prior to perfusion
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.
> From: email@example.com on behalf of
> Caroline Bass
> Sent: Wednesday, February 1, 2006 10:52 AM
> To: Histonet
> Subject: [Histonet] collecting fresh tissue prior to perfusion
> 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.
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