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. 2020 Oct 29;10:18561. doi: 10.1038/s41598-020-75517-4

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Impact of ligature-induced PD on plaque-associated microglia (PAMs) in 5xFAD mice. Fluorescent photomicrographs containing Iba1+ microglia (red), thioflavin S (ThS) dense-core plaques (green), and DAPI cell nuclei (blue) were taken of cortical layers 4–5 (CTX) from Non-Lig and Lig 5xFAD mice (×20 objective, 2 images/section, 3 sections/animal). Each fluorescent channel was automatically thresholded; an ROI around each ThS+ plaque (including a 6.5 μm buffer-zone) was generated; and channels were analyzed within and without the ROI. (A) Dense-core plaque densitometry (%ThS+), microglia densitometry (%Iba1+), and the percentage of PAMs (%Iba1+ staining within the immediate proximity of ThS+ plaques), *p < 0.05. (B) Representative fluorescent photomicrographs of Non-Lig (left panels) and Lig (right panels) 5xFAD mouse cortex. Scale bar = 100 μm. Bottom row: Iba1, ThS, DAPI, and merged channels from a representative image of a PAM.