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TTC staining

YC Yun-An Chen
YT Yi-Chieh Tsai
YC Yi-Dao Chen
DL Der-Zen Liu
TY Tai-Horng Young
LT Li-Kai Tsai
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Rats were decapitated at day 15 after MCAO and the brains were removed quickly. The sections at 2 mm thickness were cut using adult rat brain slicers (Zivic Instruments, Pittsburgh, PA) and stained with 2% solution of 2,3,5-triphenyltetrazoliumchloride (TTC) (Sigma-Aldrich) at 37 °C for 10 min to analyze the infarcted volume. The brain regions were quantified using Image J (National Institutes of Health, Bethesda, MD). The infarct area in TTC staining of each slice was obtained by subtracting the normal ipsilateral area from that of the contralateral hemisphere to reduce overestimation of the infarct area resulting from edema in the acute stage of ischemia (N = 5 for each group). We calculated the percentage of infarcted volume using the formula: Infarcted volume (%) ={[the intact contralateral hemisphere volume − (the ipsilateral hemisphere volume − the infarcted volume)]/the intact contralateral hemisphere volume} × 100%23.

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