Re: Perfusion Setup

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In a message dated 98-10-20 11:13:46 EDT, writes:

<< I need to set up a well ventilated area in which we can work
 and also have a way to collect the run off of the chemicals.

 If anyone else is perfusing please tell me what you have
 found to work. It can be a purchased unit or something that
 has been self designed.

Hi Clover,

The system I have used is definiely low-tech, but it works well.  Do you have
access to a fume hood?  If so, you can place in the hood a shallow aluminum or
glass baking dish to catch the runoff.  Cut a piece of styrofoam that is
smaller than the inside of the dish so it fits inside, and use the styrofoam
to pin out the animal.  You can hang intravenous bags with the pre-rinse and
fixative on the outside of the fume hood, and use intravenous tubing sets to
deliver the reagents after the vessel of choice is cannulated.

You will probably want to hang the I.V. bags about 80cm - 1m above the level
of the dissecting set-up so you will have a sufficient pressure head to
deliver the reagents.

Just make sure the tubing sets have a "piggyback" fitting on them.  For
example, if you pre-rinse with a buffer containing maybe a little heparin in
it, you can then "piggyback" the second fixative line to deliver the fixative
without removing the first needle.

The blood volume of a rat is so small that you won't have to worry about
spill-over.  Most animals of this size perfuse quite well with about 50ml of
pre-rinse and 100ml of fixative.  If you perfuse the whole animal you'll know
when it's fixed, because the head and the tail will be quite stiff.

If you don't have access to a fume hood, try and get a small bench-top hood
(one with a fan and a charcoal filter in it).  One thing is for certain, you
will need to somehow filter or vent the fumes from the fixative!

Good luck,

Bob Chiovetti

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