Animal is deeply anesthetized or euthanized. Reasonable anesthesia
is intraperitoneal pentobarbital 100 mg/kg after appropriate
restraint and/or neurolepsis with ketamine. Animal must be in
deepest level of anesthesia for appropriate ablation of pain and
vascular reactivity. After appropriate immobilization and confirmation
of anesthetic plane by corneal reflex, breathing pattern, and
lack of pain response, chest wall is rapidly opened within 5 to 15
s. Heart should still be beating. Chest wall should be excised or
gently reflected without torquing great vessels or carotids, and
attention given to avoiding damage to vessels. Rapid, but careful
technique is sufficient for most experiments; gold standard is total
control of respiration and blood pressure through artificial ventilation
and vascular access.