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. 2023 Sep 14;8(10):1834–1845. doi: 10.1038/s41564-023-01469-w

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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. 1 — a–d Negative stain EM of a a dormant spore (S), b a germinated spore with polar tube (PT) extended, c the tip of a PT with sporoplasm (*) passing through at high magnification and d a germinated spore post transfer, with sporoplasm egressing terminally (*). e Bright field light microscopy showing that microsporidia can be germinated on holy carbon (Quantifoil) grids. PTs are indicated with blue arrowheads. f-h CryoEM of microsporidia germinated on Multi-A Quantifoil grids, showing a PT emerging from a spore at three different magnifications. The PTs span Quantifoil holes, which is an important requirement for cryoET. g, h Higher magnification images of dashed area in f. h The PT is confined by a membrane (M) and proteinaceous PT wall (W). Transported content is visible. (R) ribosomes, (V) vesicle and (DM) double membrane. These images are representative of hundreds of micrographs collected from multiple (5) sample preparations.