Protocol Text Before proceeding to surgery, anesthetize a male mouse (C57BL/6) that is 6 weeks of age by inhalation of 2-5% isoflurane.
{"action": "anesthetize", "output": "", "reagent": ["isoflurane"]}
Perform a toe pinch to confirm that the animal is sufficiently anesthetized.

The unconscious mouse is placed on its back on a warm pad to maintain a constant temperature of 37 °C. #####%%%%% output "isoflurane-filled" are used in line10
{"action": "put", "output": "isoflurane-filled", "temperature": ["37 \u00b0C"]}
The precordial chest on the left side is shaved.

Secure the mouse to an intubation device with an isoflurane-filled chamber.
{"action": "secure", "output": "", "device": ["an intubation device"], "reagent": ["isoflurane-filled"]}
Insert a tube of 0.2 inner diameter into the mouse's trachea through mouth, then place the intubated mouse on an aseptic surgical area (37 °C by a temperature controller) ); . #####%%%%% output "ophthalmic ointment" are used in line16
{"action": "insert", "output": "ophthalmic ointment", "container": ["the mouse's trachea", "mouth"], "device": ["a temperature controller"]}
disinfect the area with betadine

Apply ophthalmic ointment to the mouse's eyes.
{"action": "apply", "output": "", "reagent": ["ophthalmic ointment"]}
Use sterile scissors to make a 1.5 cm incision along the mid-axillary line.
{"action": "use", "output": "", "length": ["the mid-axillary line"]}
Scissors are used rather than a scalpel to avoid injury to the underlying tissue in mice. #####%%%%% output "injury" are not used
{"action": "use", "output": "injury", "reagent": [""]}
Then, use blunt-tip vessel scissors to dissect .

Perform a left thoracotomy between the third ribs to visualize the anterior surface of the heart retract the thoracic wall using a suitable retractor in order to improve visualization .

Remove the pericardium by using toothed forceps .
{"action": "remove", "output": "", "device": ["toothed forceps"]}
Ligate the left coronary artery by placing the needle (attached with 7-0 ethilon suture) beneath the artery with a band of myocardium between the ligature .

A successful occlusion of coronary artery is verified by hypokinesis/akinesis of the anterior left ventricular wall e.g.
{"action": "verify", "output": "", "device": [""]}
ST segment elevation, QRS broadening Close the chest by placing two stitches (6-0 ethilon suture) on the third ribs. #####%%%%% output "ethilon suture" are used in line34
{"action": "close", "output": "ethilon suture", "container": ["the chest", "the third ribs"]}
Use 5-0 ethilon suture to close the skin. #####%%%%% output "isoflurane anesthesia" are used in line38
{"action": "use", "output": "isoflurane anesthesia", "reagent": ["ethilon suture"]}
For post-operative analgesia, administer a single intraperitoneal injection of buprenorphine (0.05 mg/kg).

The animals will spontaneously recover within 2-5 min after stopping isoflurane anesthesia. #####%%%%% output "buprenorphine" are used in line42
{"action": "recover", "output": "buprenorphine", "reagent": ["isoflurane anesthesia"]}
Finally, place the mouse in a warm cage .

provide 0.6 mg/ml of buprenorphine in the drinking water to prevent post-operative distress #####%%%%% output "post-operative distress" are not used
{"action": "provide", "output": "post-operative distress", "reagent": ["buprenorphine"]}
Closely monitor the mouse for the first 24 hours after surgery.

Echocardiography is performed to evaluate cardiac function 3 days after myocardial infarction. #####%%%%% output "cardiac function" are used in line52
{"action": "perform", "output": "cardiac function", "device": [""]}
Successful procedure is verified by akinesia in the anterior left ventricular wall (see the enclosed video in Results).

At the end of the experiment, the heart will be collected .

stored in -80 °C for further investigations
{"action": "use", "output": "", "temperature": ["-80 \u00b0C"], "mass": [""], "reagent": ["cardiac function"]}
Please recommend JoVE to your librarian.
