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. 2021 Sep 5;227(1):23–47. doi: 10.1007/s00429-021-02373-x

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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/.

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Fig. 17 — a, b, Pictures taken during the preparation procedure for cutting the midbrain specimen. The specimen is placed in a Perspex box (20 × 20 × 20 cm), with a titanium base and four mountable/ removable walls. b Approximately 2 L of a 5% agar solution was poured into the box, and when it started to set, the brain was placed on the agar base and the anterior commissure—posterior commissure was horizontally aligned; c the hardened agar block is placed in a box with rails of 5 mm spacing. d–f Examples of ACPC-parallel cuts of 5 mm thickness (macro, unstained)