Light microscopy allows for the inexpensive and fast detection of neuronal /glial cell demise and estimation of infarct and traumatic lesion volumes ; the direct correlates of cell death . Quantitative assessment of brain tissue damage following stroke , traumatic brain injury (TBI ) or neurodegenerative diseases , and recovery after therapeutic intervention has been facilitated by recent developments in computer-assisted image analysis technologies that enable more objective and accurate morphometric quantification of cell injury in whole brain sections . In this chapter, the proposed workflow describes what tasks need to be fulfilled to visualize and gauge cell death characterization by histological stains and immunohistochemical markers .
Artifact-free brainfixation/processing into cryo- or paraffin blocks Digital pathology TBI neuropathology and hemorrhagescores IHC Algorithm-based morphometry of death events
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This work was supported by the National Institutes of Health (NIH) Grant NS-036821 and the Department of Defense (DoD) Grant W81XWH-10-1-0847 awarded to Stan Krajewski. We are greatly indebted to Dr. Allen Olsen from Aperio Technology, Inc. for collaborative development and beta-testing of Cell/Cytoplasmic v2 algorithm. We thank Xianshu Huang for high-quality histological preparations and immunocytochemistry, Tomi Omel and Francisco Beltran for excellent animal care assistance.
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