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. 2021 Jan 28;18:33. doi: 10.1186/s12974-021-02093-3

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© The Author(s) 2021

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. 1 — Microglia-specific ChETA expression in organotypic hippocampal slice cultures. a Schematic overview of microglia-specific expression. The microglia-driver line Cx3cr1-CreERT2 (blue) is crossed with a reporter mouse line (R26-LSL-tdTomato, red) and the ChETA mouse line (R26-LSL-ChETA, green). After injection of (Z)-4-hydroxytamoxifen, the tdTomato and ChETA are expressed in microglia. b Illustration of the Channelrhodopsin-variant ChETA activated by blue light. Scale bar 25 μm. c Immunostaining using antibodies against the reporter (tdTomato - red) and microglia (iba1 - cyan). d Graphic illustration of the hippocampal structure and the investigated area for microglia morphology in e (red square). e Z-projection of confocal images acquired for Sholl analysis of microglia at 3, 13, 29 DIV, and in vivo. Scale bar: 25 μm. f Confocal image of a microglia cell in organotypic slice culture which was fixed with PFA and stained against the microglia marker iba1. Overlay with IMARIS analysis (magenta). g Result of microglia branch detection with color coding by branching level. h Sholl analysis of microglia over time (number of intersections versus distance from cell body). i Quantification of % microglia cells between dentate gyrus and CA1 relative to total cell count (DAPI)