Histological Quantitation of Brain Injury Using Whole Slide Imaging: A Pilot Validation Study in Mice

Abstract

Quantitative assessment of serial brain sections provides an objective measure of neurological events at cellular and molecular levels but is difficult to implement in experimental neuroscience laboratories because of variation from person-to-person and the time required for analysis. Whole slide imaging (WSI) technology, recently introduced for pathological diagnoses, offers an electronic environment and a variety of computational tools for performing high-throughput histological analysis and managing the associated information. In our study, we applied various algorithms to quantify histologic changes associated with brain injury and compared the results to manual assessment. WSI showed a high degree of concordance with manual quantitation by Pearson correlation and strong agreement using Bland-Altman plots in: (i) cortical necrosis in cresyl-violet-stained brain sections of mice after focal cerebral ischemia; (ii) intracerebral hemorrhage in ischemic mouse brains for automated annotation of the small regions, rather than whole hemisphere of the tissue sections; (iii) Iba1-immunoreactive cell density in the adjacent and remote brain regions of mice subject to controlled cortical impact (CCI); and (iv) neuronal degeneration by silver staining after CCI. These results show that WSI, when appropriately applied and carefully validated, is a highly efficient and unbiased tool to locate and identify neuropathological features, delineate affected regions and histologically quantify these events.

Combined Diffusion Tensor Imaging and Quantitative Susceptibility Mapping Discern Discrete Facets of White Matter Pathology Post-injury in the Rodent Brain

“…In the step (i) and (ii) of Figure 6, we demonstrated that the axons that were normal before injury appear to be swollen within 5 h post-injury that has stretched the myelin sheath reflected by increase the radial diffusivity (perpendicular) without any change in the axial (parallel) diffusivity (82) (Figure 1). This appeared to be followed by ischemia and inflammation within days leading to the shrinkage and tearing of the axonal bundles (83,84) leading to the reduction of both axial and radial diffusivity (3,68,69) noticed at 3 days post-injury in our study (Figure 2). These pathologies further progress to myelin fragmentation/loss inflammation that may further increase radial diffusivity susceptibility (see Figures 1, 3 for RD and susceptibility increase and Figures 4, 5 for myelin loss and gliosis.…”

Sigma-1 Receptor Modulates Neuroinflammation After Traumatic Brain Injury

“…A previously published wire hanging test was used in this test (Chen et al 2014). 55 cm above the table, there is metallic wire between two posts.…”

Development and validation of an app-based cell counter for use in the clinical laboratory setting

“…are familiar to pathologists and laboratorians alike, the approach to demonstrating similarity between data sets is often less well-defined. Some studies rely exclusively on the correlation coefficient to establish statistical similarity; however,[ 12 13 14 15 16 17 ] the combination of the correlation coefficient with Bland–Altman analysis is a more stringent approach to demonstrating functional equivalence between a novel and an established method and is subject to fewer sources of error. [ 9 10 12 13 16 ] The superiority of a combined approach is evidenced by our results for peripheral blood basophils: Although only moderate correlation was observed for this cell category ( r = 0.684; Figure 2f , upper panel), the accompanying Bland–Altman analysis [ Figure 2f , lower panel] demonstrated that this lack of correlation does not represent a systematic discrepancy between the two methods.…”