Both injured and sham control swine were euthanized at the post injury time points noted above as previously described [32 (link), 33 (link)]. Briefly, transcardial perfusion was performed under surgical plane anesthesia using heparinized saline (2L) followed by 10 % neutral buffered formalin (NBF) (8L). The brain was removed, extracted, and post-fixed in 10% NBF for 1 week. Thereafter, the brains were sectioned at 5mm in the coronal plane, with tissue sections processed for standard paraffin embedding in an automated tissue processor (Shandon Scientific Instruments, Pittsburgh, PA). Serial sections (8 μm) were cut on a Leitz rotary microtome (Leica, Malvern, PA) then mounted on Fisherbrand Superfrost/Plus microscope slides (Fisher Scientific, Pittsburgh).
For human brain tissue preparation, the intact brain was immersed in 10% formol saline at autopsy and fixed for at least 3-weeks prior to dissection. Sampling was achieved using a standardized protocol and paraffin embedding was performed as described previously [24 (link)]. The region of cingulate gyrus including corpus callosum was selected in this study given its midline location containing white matter and known susceptibility to injury [1 (link), 2 (link), 29 (link), 31 (link), 32 (link), 36 (link)].
For human brain tissue preparation, the intact brain was immersed in 10% formol saline at autopsy and fixed for at least 3-weeks prior to dissection. Sampling was achieved using a standardized protocol and paraffin embedding was performed as described previously [24 (link)]. The region of cingulate gyrus including corpus callosum was selected in this study given its midline location containing white matter and known susceptibility to injury [1 (link), 2 (link), 29 (link), 31 (link), 32 (link), 36 (link)].