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. 2023 Jul 10;20(8):1256–1265. doi: 10.1038/s41592-023-01936-6

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© The Author(s) 2023

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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Extended Data Fig. 6 — Corresponding orthogonal planes in xy- and xz-directions from 6 consecutive LIONESS measurements of the same volume in the neuropil of an organotypic hippocampal slice culture. The volume was initially imaged 4 times within one hour and then again after one day and after three days. Magnified views: Subregion with dendritic spines revealing morphodynamics. Scale bars, overview: 2 µm, magnified views: 500 nm. White arrowheads at image edges indicate the position of corresponding orthogonal planes. Maximum intensity projections spanning 150 nm. Additional dark regions on day 1 and day 3 likely represent branched processes of a damaged cell that took up dye after repeated manual mounting of the sample (supported by a membrane for interface tissue culture), transfer to the microscope, volumetric imaging, unmounting, and transfer back to the tissue culture incubator. The specific measurement sequence applied here was done for n = 1 sample.