How to perform cardiac perfusions - (Aug/23/2006 )

Hello,
Does anyone have protocols on performing cardiac perfusion in rats?
Need it to fix brain tissue for immunohistochemistry purposes.
What materials/equipment do I need, how do I set it up?

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Hello!

I don't have any established protocol for cardiac perfusion in rats, but what do you think about this:
- to open the chest and discard the sternum and ventral part of the ribs (after having anaesthetized the rat!)

- and to inject the fixer solution (for instance, paraformaldehyde 4%, with or without glutaraldehyde) in the left ventricle.

I know some people work like this to fix abdominal organs in mice. I think it may work also to fix brain.

A simple way to check if the injected solution went where you wanted is to inject (instead of the fixer solution) an Evan's blue (or other blue colouring with similar molecular weight) solution (so, to check if your perfusion protocol is ok): if the organ you want to target get a "blue coloration", you know it's successful! Then, just do the same with the fixer solution (with another rat).

If it doesn't work with injection in the left ventricule, perharps try to inject directly in the carotids arteries...

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