Assessing Infarct Volume in Stroke Mice

The infarct volume was assessed at 6 h and 1–14 days after PTS. Mice were deeply anesthetized with sodium pentobarbital (50 mg/kg), and their brain tissues were collected and stored in a −20 °C refrigerator for 20 min before being sliced into sections that were 2 mm thick in the coronal plane. The brain sections were then stained with 2% TTC (Sigma-Aldrich) for 30 min at 37 °C in the dark, with the slices being turned evenly every 8 min to ensure even contact with the TTC staining solution. Subsequently, the tissues were fixed in a 4% paraformaldehyde (PFA) solution to ensure optimal tissue hardness. After staining, the infarct appeared white, while the normal brain tissue was stained red. We measured the infarct area using Image J (Version 2.0.0) as described [17 (link),18 (link)], and calculated the brain infarct volume percentage (BIVP).