| | Re: mouse hepatocytes isolation
Yes, practice perfusion through the portal vein; this works the best. You will miss the vein and/or poke all the way through the vein several times before you get it right but after several attepmts you wil succeed. It took me many mice before I got good at it.
Use a new, sharp needle for each mouse for best results.
As soon as you insert into the portal vein, turn on the pump, and snip the vena cava, the liver will turn pale.
I first perfuse with a anti-coagulant perfusion buffer to clear the blood, and then perfuse with collagenase. Saline solution will work for practice, but it is not an anti -coagulant.