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Begin with an anesthetized mouse in the prone position, with its head shaved.
Make an incision in the skin over the head.
Drill a small hole in the skull, stopping just before the brain layer or dura mater to avoid damaging brain tissue.
Use blunt forceps to penetrate the final tissue bridge and remove any sharp edges.
Insert a balloon catheter through this opening, ensuring it is fully within the cranial cavity. This catheter is prefilled with saline and free of air.
Infuse additional saline through the catheter to gradually inflate the balloon, increasing intracranial pressure.
As the pressure rises, it compresses the brain's blood vessels, restricting blood flow and oxygen to neurons, leading to neuronal death.
This causes the mouse's tail to stiffen and breathing to stop, indicating brain death.
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