Transient Focal Ischemic Stroke Model

Seventeen male C57Bl6/J mice (Janvier, France, weight range 20–25 g, age range 8–10 weeks) were used. Anesthesia was initiated by using 3% isoflurane (Abbott, Cham, Switzerland) in a mixture of O₂ (200 ml/min) and air (800 ml/min) and maintained with 1.5–2% isoflurane. Prior to surgery, a local analgesic (lidocaine, 0.5%, 7 mg/kg) was administered subcutaneously. Temperature was controlled during the surgery and kept constant at 36.5 ± 0.5 °C with a feedback-controlled heating pad system. The surgical procedure was carried out as described [30 (link), 31 (link)]. The middle cerebral artery was occluded for 1 h. After surgery, buprenorphine was administered as subcutaneous injection every 6–8 h on the day of surgery (Temgesic, 0.1 mg/kg b.w) and supplied thereafter via the drinking water (1 mg/kg) for 36 h.
tMCAO animals were assessed with MRI at 2 h (n = 3), 4 h (n = 4), 6 h (n = 3), 12 h (n = 4), 24 h (n = 3), and 48 h (n = 6) after reperfusion, with the majority of animals being measured at two time points. ADC maps of all investigated mice were inspected. Animals were analyzed when a lesion was present on the ADC maps.