2.7. Transient middle cerebral artery occlusion (tMCAO) in mice

Transient MCAO surgery was performed as described previously.²⁵, ²⁶ Mice were anesthetized with isoflurane (1.5‐2%) and oxygen/nitrous oxide (30%/70%). Firstly, an incision was made in the middle of the neck, and the external carotid artery, internal carotid artery and left common carotid artery were isolated carefully by microscopic curved tweezers. A 6‐0 nylon monofilament suture (Dermalon, 1756‐31, Covidien) coated with silicon was gently inserted from the external carotid artery to the internal carotid artery. The insertion stopped until it blocked the middle cerebral artery. The insertion depth was about 0.8‐1 cm. After 90 minutes of ischemia, the suture was withdrawn gently to allow reperfusion. Successful occlusion and reperfusion of the MCA were confirmed by laser Doppler flowmetry (Moor Instruments). Regional blood flow declined more than 80% compared with the baseline and recovered to 80% of the baseline is considered as a successful model.