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[topino8]

HI,

I'm experiencing problems in the perfusion of mouse spleen. I'd like to separate resident splenocytes from circulating leukocytes in the spleen. I tried to use standard techniques, like perfusion with PBS from the heart (under terminal anaesthesia) or perfusion from the aorta. Big organs like lungs and liver are cleared, but the spleen remains full of blood cells (probably due to the peculiar vascular anatomy).

Has anyone noticed spleen whitening during organ perfusion with other techniques???

thanks

[gfischer]

There may be too much clot formation to get complete perfusion.  When I perfuse, I inject some heparin into the heart before perfusing with saline (I use about .3 cc for a 300g rat).  If you have solid heparin, you can also add it to the saline you use for perfusion.