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5-Triphenyltetrazolium chloride (TTC) staining
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Brains were harvested and cut into 1 mm-thick slices using a mouse brain matrix (Harvard Apparatus, MA, USA). Brain slices were incubated in a 1% 2,3,5-triphenyltetrazolium chloride (TTC) solution in phosphate buffered saline for 30 min. TTC is converted by active mitochondria to a red compound, thereby staining metabolically active tissue red and leaving infarcted tissue white. Stained slices were imaged for calculation of infarct volume in Image J (National Institutes of Health, Bethesda, MD).
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