2,3,5-Triphenyltetrazolium Chloride (TTC) Staining

We prepared 1% TTC (T8877-25G; Sigma, USA; W/V) with PBS, and dissolved the mixture at 37°C. The brain tissue section was incubated with 10 ml TTC solution at 37°C for 10 min to stain the tissue evenly. The normal brain tissue was stained bright red while the infarct area was stained pale white. The whole-infarct volumes and whole-brain volumes were determined via Image-Pro Plus 6.0 analysis software. The infarct volumes were calculated according to the ratio of the whole-infarct volumes to the whole-brain volumes.