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Fig. 5 | Acta Neuropathologica Communications

Fig. 5

From: Identification of a dysfunctional microglial population in human Alzheimer’s disease cortex using novel single-cell histology image analysis

Fig. 5

Iba1low cells are not spatially distributed around Aβ plaques in AD cortex. To characterise the spatial distribution of Iba1low cells relative to Aβ plaques, AD middle temporal gyrus sections were subjected to two rounds of immunofluorescent staining; (1) Iba1 was immunofluorescently co-labelled with L-Ferritin and HLA-DR, and (2) amyloid beta and tau. The images from the two rounds of staining were aligned, allowing us to determine the spatial localisation of Iba1-L-Ferritin-HLA-DR cells relative to amyloid beta plaques and tau tangles. Iba1-L-Ferritin-HLA-DR cells were identified near amyloid beta plaques but showed no spatial distribution around tau (a). A master mask of total Iba1-L-Ferritin-HLA-DR cells was generated. The location of Iba1-L-Ferritin-HLA-DR cells was classified as plaque, plaque-adjacent, or non-plaque (b). The proportion of total Iba1-L-Ferritin-HLA-DR cells at plaque, plaque-adjacent, and non-plaque spatial locations was quantified (c); data shown as mean ± SD (n = 8). The Iba1low population was gated out from total Iba1-L-Ferritin-HLA-DR cells (as shown in b) and no difference in the abundance of Iba1low cells at plaque, each 5 µm plaque-adjacent interval, and non-plaque spatial locations was identified (d); data shown as median percentage of total Iba1-L-Ferritin-HLA-DR cells at each spatial location (n = 8). When the 5–50 µm intervals were grouped into plaque-adjacent, the abundance of Iba1low cells at plaque, plaque-adjacent, and non-plaque spatial locations remained unchanged (e); data shown as the percentage of total Iba1-L-Ferritin-HLA-DR cells at each spatial location with a single point per case (n = 8). The abundance of Iba1low cells was compared between amyloid beta spatial locations using a repeated measures two-way ANOVA with the Geisser–Greenhouse correction and Tukey’s multiple comparisons test. amyloid beta

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