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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. 3 — a) A pollen pellet composed of many individual pollen grains. b) Maximum projection of a single-photon light-sheet (mesoSPIM) stack of the sample. As the sample was illuminated from the side, only part of the pollen pellet is visible. c) Maximum projection of a stack taken in air after transferring the sample into the Schmidt objective. d)–e) Maximum projections of stacks taken at higher zoom. f) The immersion chamber was then filled with ethyl cinnamate (ECI) with an index of 1.56. This has several consequences: Firstly, at the same zoom level, the FOV shrinks by 1.56× as in the Schmidt objective, the product NA × FOV is constant. Secondly, the pollen are now much better index-matched and the microscope can also start visualizing the interior of the pollen grains. Therefore, the visual appearance of pollen grains is different than in air. g)–h) Zoomed-in maximum projections. Similar sample regions in both images are highlighted with arrows. The experiment was repeated once.