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. 2022 Mar 31;23:21. doi: 10.1186/s12868-022-00703-1

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© The Author(s) 2022

Open AccessThis 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — Imaging of autofluorescent granules in microglia on live slices and in vivo. A Diagram illustrates the strategy to image calcium signals in hippocampus CA1 microglia cells on live brain slices; B A representative Z-projected image of 60 GCaMP6s frames during 15 min imaging of a microglia cell on Cx3cr1^CreER: GCaMP6s mouse brain slice. The arrow points to the soma of a microglia cell, which contains an autofluorescent granule. ROI1 circles the soma of microglia, excluding the autofluorescent granule, while ROI2 including it. Right top: the GCaMP6s image in basal condition. Right bottom: GCaMP6s image during application of 100 µM UDP. C F/F₀ in ROI1 and ROI2 during 15 min imaging, at 5 min of which 100 µM UDP was applied. D The peak of F/F₀ of GCaMP6s signals during UDP application in ROIs either excluding (ROI1) or including (ROI2) the autofluorescent granules (n = 5 cells from 3 mice, paired t-test). E Emission spectrum of the autofluorescent puncta as in panel (B) by spectral imaging (n = 5 cells from 3 mice). F Representative images of a microglia from Cx3cr1^GFP mice with GFP and autofluorescence simultaneously imaged by 2-photon microscopy through cranial window. The white arrows point to the autofluorescent granule in different time points. Bars represent means ± SD