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. 2018 Aug 27;9:3460. doi: 10.1038/s41467-018-05976-x

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© The Author(s) 2018

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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Fig. 1 — Correlative light and electron microscopy (CLEM) for visualizing wild-type V. cholerae inside the contractile vacuole (CV). a Low- and high-resolution imaging of an infected amoeba. GFP-tagged V. cholerae were seen to be localized inside of a CV of A. castellanii using confocal laser scanning microscopy (CLSM; low resolution) in fixed samples. Shown are a merged image of the transmitted light channel and the green channel (left) and the green channel image alone (second from left). After staining of the sample, the same amoeba was imaged at high resolution using transmission electron microscopy (TEM; right images). Scale bar in all images: 5 µm. b 3D reconstruction of the colonized CV. The region containing the amoeba shown in a was serially thin sectioned (50 nm thickness) and serial images were taken with the TEM. These images were then aligned to generate a 3D model of the colonized amoeba. Shown are snapshots of the resulting 3D reconstruction movie (Supplementary Movie 1)