Tissue Processing

After completion of live experiments, anesthetize the mouse with institution-approved substance at correct dosage.

Ensure the mouse is deeply anesthetized via the toe-pinch test and that there is no reaction. Place the mouse with its chest facing up above a basin to capture blood, and secure the paws so that the body doesn't move.

While there is still a heart beat, open chest cavity using small scissors and forceps, making sure not to puncture organs.

With a small needle/syringe attached to a pump, pierce the left (on your right side) ventricle. Pierce the left atrium to allow blood to flow through.

Perfuse the mouse with 1x PBS to clear the blood.

Once blood has been cleared, perfuse the mouse with 4% PFA.

Once the whole body is perfused, dissect all needed tissues and organs.

Post-fix the dissected tissues in 4% PFA for 4-6 hours.

After post-fixation, gently dry the tissue.

Embed each tissue to OCT block.

Section tissue at 40um thickness using cryostat.