MAP2-stained sections were analyzed to quantify grey matter injury [32–34 (link)]. For quantification of infarct size, every 50th section throughout the brain was stained for MAP2. Images were captured using an Olympus light microscope with 1.25× magnification and analyzed using ImageJ (version 1.51j8, ImageJ.nih.gov/ij">http:/ImageJ.nih.gov/ij , National Institutes of Health, U.S.A.). Infarct size was calculated by subtracting the ipsilateral MAP2-stained area from the contralateral and expressing the difference as percentage of the stained contralateral area [32 (link)]. Quantifications were made for the whole hemisphere and for cortex specifically. All quantifications were performed in a blinded manner.
Iba-1 stained sections were analyzed by phenotype scoring, using a modified version of a previously described scoring method [35 (link),36 (link)]. Briefly, affected brain regions (striatum, upper and lower cortex, cerebral peduncle, globus pallidus and substantia nigra) were given a score 0-4, based on microglial phenotype and accumulation (Supplementary Figure S1). Each region was scored using a light microscope (Olympus) with three 20× magnified pictures at each level. A mean score for each region was calculated at each level, and then a mean for the region throughout the different levels in which the region was present.
Iba-1 stained sections were analyzed by phenotype scoring, using a modified version of a previously described scoring method [35 (link),36 (link)]. Briefly, affected brain regions (striatum, upper and lower cortex, cerebral peduncle, globus pallidus and substantia nigra) were given a score 0-4, based on microglial phenotype and accumulation (Supplementary Figure S1). Each region was scored using a light microscope (Olympus) with three 20× magnified pictures at each level. A mean score for each region was calculated at each level, and then a mean for the region throughout the different levels in which the region was present.
Free full text: Click here
