Scanjet 2c

Under anesthesia the rat was decapitated using a rodent guillotine, the brain was dissected out and cooled in ice water for 5 minutes and sliced in 2 mm slices using a rat brain matrix (RBM-4000, ASI Instrument Inc., USA). Subsequently, the slices were soaked in 2% 2, 3, 5-triphenyltetrazolium hydrochloride solution (TTC; Sigma-Aldrich Sweden AB, CAS# 298–96–4, Stockholm, Sweden) at 37 °C for 15 minutes and scanned (ScanJet 2c, Hewlett-Packard, Palo Alto, CA, USA). Infarcts were measured in a similar way as described by Goldlust et al.43 (link), with an automatic 40% green spectrum threshold (SigmaScan Pro 5, Systat Software Inc, San Jose, CA, USA). Edema correction was performed according to the following formula:
Corrected infarct volume, as part of one hemisphere = [crude infarct area × [contralateral hemisphere/ipsilateral hemisphere]]/contralateral hemisphere

Under anesthesia (see “Surgical procedures” above) the rat was decapitated using a rodent guillotine. The brain was dissected out, cooled in ice water for 5 min and subsequently sliced in 2 mm slices using a rat brain matrix (RBM-4000, ASI Instrument Inc., USA). The slices were then soaked for 15 min in 2% 2,3,5-triphenyltetrazolium hydrochloride solution (TTC; Sigma-Aldrich Sweden AB, CAS# 298-96-4, Stockholm, Sweden) at 37°C and scanned (ScanJet 2c, Hewlett-Packard, Palo Alto, CA, USA). Infarcts were measured in a similar way as described by Goldlust et al. (8784144), with an automatic 40% green spectrum threshold (SigmaScan Pro 5, Systat Software Inc, San Jose, CA, USA). Total infarct volume was calculated indirectly according to Swanson et al. [48 (link)]:
Corrected infarct volume = [contralateral hemisphere − [ipsilateral hemisphere − crude infarct]]/contralateral hemisphere.
The indirect method (calculation based on remaining viable tissue in the lesioned hemisphere) was chosen to minimize error introduced by loss of brain tissue due to the infarction 7 days after MCAo. Direct measurement of infarct size (pixels) was used to calculate proportion of infarct in cortex and striatum, respectively.

24 h after MCAo, the rat was anesthetized with Isoflurane (Forene^®, Abbott Scandinavia AB, Solna, Sweden), 4.5% delivered in a 30/70 mixture of O₂/N₂O, and decapitated using a rodent guillotine. The brain was dissected out, cooled in ice water for 5 min and sliced in 2 mm slices using a rat brain matrix (RBM-4000, ASI Instrument Inc., USA). The slices were subsequently soaked in 2% triphenyl tetrazolium chloride solution (TTC; Sigma-Aldrich Sweden AB, CAS# 298-96-4, Stockholm, Sweden) at 37 °C for 15 min and scanned (ScanJet 2c, Hewlett-Packard, Palo Alto, CA, USA). An example of TTC stained brain slices are provided in Fig. 5. Infarcts were measured at large according to the method described by Goldlust et al.’s [50 (link)], with an automatic 40% green spectrum threshold (SigmaScan Pro 5, Systat Software Inc, San Jose, CA, USA). Edema correction was performed according to the following formula:Fig. 5

Example of scanned brain slices from a rat subjected to 60 min of middle cerebral artery occlusion. The brains were sliced in 2 mm slices that were soaked in 2% triphenyl tetrazolium chloride solution (TTC) for 15 min and subsequently scanned

$$\text{Corrected}\text{infarct}\text{volume},\text{as}\text{part}\text{of}\text{one}\text{hemisphere}=\left[\text{crude}\text{infarct}\text{area}\times \left[\text{contralateral}\text{hemisphere/ipsilateral}\text{hemisphere}\right]\right]/\text{contralateral}\text{hemisphere}$$