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. 2018 Sep 27;8:14491. doi: 10.1038/s41598-018-32820-5

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© The Author(s) 2018

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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3D-SEM reconstruction of the primary apical dendrite of single AVC-labeled pyramidal neuron and its subcellular features such as mitochondria. (A) Montages of serial electron micrographs including the DAB labeled pyramidal neuron shown in Fig. 1D,E. (see Supplemental Movie 3). (B,C) 3D reconstruction of the plasma membrane (B) and the mitochondria (C) of its labeled apical dendrite (see Supplemental Movies 4 and 5). To find out the region of interest, 256 serial sections were imaged with 30 nm/px resolution. The region of interest was re-imaged with 3 nm/px resolution through 103 serial sections and reconstructed. (D) Parameters tested to optimize the DAB staining and preservation of the membrane structures for SSEM. Bold indicates the optimal conditions used in the paper.