4.4. TTC Staining

The infarct volume was measured using TTC, a standard method for the fast and reliable delineation of cerebral ischemic injury [60]. Twenty-four hours after the onset of transient ischemia, the mice were euthanized and the intact brains were rapidly removed and washed in PBS (pH 7.4). Brain tissue from an area 4 mm anterior and 5 mm posterior to the bregma was cut into serial 1 mm thick slices using a mouse brain slicer. The sliced brain tissue samples were stained with 2% TTC (Sigma-Aldrich, St. Louis, MO, USA) for 30 min at 37 °C in the dark, followed by overnight immersion in 4% paraformaldehyde (PFA) in 0.1 M PBS at pH 7.4 and 4 °C [61]. Images of TTC-stained brain sections were obtained using a digital camera.