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

Jacob M. Stillman, Francisco Mendes Lopes, Jing-Ping Lin, Kevin Hu, Daniel S. Reich and Dorothy P. Schafer ()
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Jacob M. Stillman: University of Massachusetts Chan Medical School
Francisco Mendes Lopes: University of Massachusetts Chan Medical School
Jing-Ping Lin: Translational Neuroradiology Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health
Kevin Hu: Translational Neuroradiology Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health
Daniel S. Reich: Translational Neuroradiology Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health
Dorothy P. Schafer: University of Massachusetts Chan Medical School

Nature Communications, 2023, vol. 14, issue 1, 1-11

Abstract: 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.

Date: 2023
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DOI: 10.1038/s41467-023-42809-y

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