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Lipofuscin-like autofluorescence within microglia and its impact on studying microglial engulfment

Stillman, Jacob M
Mendes Lopes, Francisco
Lin, Jing-Ping
Hu, Kevin
Reich, Daniel S
Schafer, Dorothy P
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Abstract

Engulfment of cellular material and proteins is a key function for microglia, a resident macrophage of the central nervous system (CNS). Among the techniques used to measure microglial engulfment, confocal light microscopy has been used the most extensively. Here, we show that autofluorescence (AF) likely due to lipofuscin (lipo-AF) and typically associated with aging, can also be detected within microglial lysosomes in the young mouse brain by light microscopy. This lipo-AF signal accumulates first within microglia and it occurs earliest in white versus gray matter. Importantly, in gray matter, lipo-AF signal can confound the interpretation of antibody-labeled synaptic material within microglia in young adult mice. We further show that there is an age-dependent accumulation of lipo-AF inside and outside of microglia, which is not affected by amyloid plaques. We finally implement a robust and cost-effective strategy to quench AF in mouse, marmoset, and human brain tissue.

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Stillman JM, Mendes Lopes F, Lin JP, Hu K, Reich DS, Schafer DP. Lipofuscin-like autofluorescence within microglia and its impact on studying microglial engulfment. Nat Commun. 2023 Nov 3;14(1):7060. doi: 10.1038/s41467-023-42809-y. PMID: 37923732; PMCID: PMC10624656.

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DOI
10.1038/s41467-023-42809-y
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37923732
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This article is based on a previously available preprint in bioRxiv, https://doi.org/10.1101/2023.02.28.530224

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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/. © The Author(s) 2023; Attribution 4.0 International