To begin, secure the base plate in a sideways position in front of the surgeon. After endotracheal intubation, place the anesthetized mouse on the base plate of the purpose-built ischemia reperfusion injury tool. Use tape to fix the mouse's tail and paws.
Then fix the head using the 2-0 silk suture around the upper incisors, to prevent accidental movement. Insert the rectal thermal probe to monitor the body temperature. Use an infrared heating lamp above the mouse to maintain the temperature around 37 degrees Celsius.
Position the ECG needle electrodes in all four paws to record heart rate and ECG wave form. After thoracotomy, place a rib retractor in the intercostal space, followed by a small piece of cotton on top of the lung to protect it. Next, use a tapered needle to pass a 7-0 polypropylene suture around the left anterior descending coronary artery.
Remove the small piece of cotton and verify the continued ventilation of the left lung. Remove the rib retractor and cut off the needle. Position a segment of one millimeter PE-50 tubing over a 15 millimeter segment of PE-10 tubing.
Direct both ends of the suture through the PE-10 tubing and shift the tubing with its thicker end against the heart. Apply fucidic acid gel to the exposed intercostal space and briefly obstruct the outflow of the ventilator to allow the remaining intrathoracic air to escape. Suture the pectoral muscle using a 5-0 polypropylene X stitch above and below the level of the PE tube.
Close the skin with two 5-0 polypropylene X stitches above and two below the PE tube. Next, gradually reduce the tidal volume and respiratory rate to wean the mouse off mechanical ventilation. When the mouse starts breathing carefully disconnect the ventilator tube.
