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. 2023 Mar 30;42(1):65–71. doi: 10.1038/s41587-023-01717-8

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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. 5 — A chicken embryo was stained for Neurofilament using Alexa 568 as a secondary antibody and cleared using BABB. a) Maximum projection of a single photon light-sheet (mesoSPIM) overview dataset of the sample. b) Sample (glued to a glass capillary) prior to insertion into the Schmidt objective. c) Sample (white arrow) inserted into the Schmidt objective. d) & g) Maximum projections (mesoSPIM dataset) of a region of interest chosen for high-resolution imaging. e) & h) Maximum projections of z-stacks taken at the highlighted locations in d) & g) using the Schmidt objective and two-photon excitation. f) & i) Maximum projections of stacks taken at higher zoom level showing individual axons. The white cells with strong autofluorescence are red blood cells which have not lost their nucleus yet at this developmental stage. The images are examples from one of two imaged chicken embryos.