The optical fractionator method of stereology of the stereo-investigator software
(MBF Biosciences) was used to quantify nitrotyrosine, cleaved caspase 3, and
NeuN immuno-reactive cells in the hippocampal dentate gyrus region. For all
measurements, six sections were obtained to cover the entire impact region,
−1.60 mm to −6.3 mm from Bregma, corresponding to every 12 serial sections for
each brain. For the stereological quantitation, we used a grid spacing of 75 μm
× 75 μm in the x and y-axis and guard zones of 2 μm at the top and bottom of
each section where immuno-positive cell bodies were counted. The total number of
nitrotyrosine, cleaved caspase 3, and NeuN-positive cells in the volume of
interest were automatically determined and expressed as
cells/mm3.
Image J software (NIH) was used to measure immunoblot protein band signal
intensity, the internal length, and the central width of each endothelial
nucleus.
(MBF Biosciences) was used to quantify nitrotyrosine, cleaved caspase 3, and
NeuN immuno-reactive cells in the hippocampal dentate gyrus region. For all
measurements, six sections were obtained to cover the entire impact region,
−1.60 mm to −6.3 mm from Bregma, corresponding to every 12 serial sections for
each brain. For the stereological quantitation, we used a grid spacing of 75 μm
× 75 μm in the x and y-axis and guard zones of 2 μm at the top and bottom of
each section where immuno-positive cell bodies were counted. The total number of
nitrotyrosine, cleaved caspase 3, and NeuN-positive cells in the volume of
interest were automatically determined and expressed as
cells/mm3.
Image J software (NIH) was used to measure immunoblot protein band signal
intensity, the internal length, and the central width of each endothelial
nucleus.
