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. 2020 Aug 19;10:14014. doi: 10.1038/s41598-020-70859-5

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© The Author(s) 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Measuring synaptic densities and sizes in stacks of sections obtained by FIB-SEM. (a–c) Panoramic view of electron micrographs of the stratum lacunosum moleculare (SLM), stratum radiatum (SR) and stratum oriens (SO) imaged by FIB-SEM. (d–f) 3D rendering of synaptic junctions reconstructed from the corresponding stacks of serial sections, acquired from the strata represented in (a–c). Asymmetric synaptic junctions have been represented in green and symmetric synaptic junctions in red. (g–i) The synaptic apposition surfaces (SAS, yellow) have been automatically extracted from the three-dimensionally reconstructed synaptic junctions. The number of synapses per unit volume and the surface areas of the SAS have been measured in each stack of serial sections (see Tables 1 and 2). Calibration bar in (c): 3 µm.