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. 2017 Jun 16;7:3645. doi: 10.1038/s41598-017-03900-9

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© The Author(s) 2017

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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Correlation between 2D images and volume rendering image of E10.5 mouse embryo. (a) An E10.5 mouse embryo was prestained with hematoxylin and examined using the CoMBI technique. (b) Volume rendering image was reconstructed from 503 block-face images to show the surface structures of the mouse embryo. (c,d) At the planes c and d in (b), frozen sections were collected and fluorescently labeled for nuclei, cyclin D1, and VAMP2. The block-face images are shown in the rightmost column. The sections were examined by light microscopy, and showed nuclei prestained with hematoxylin. The sections were also examined by fluorescence microscopy, and showed nuclei labeled with DAPI. Distributions of VAMP2 (a marker of synaptic vesicles, green) and cyclin D1 (cell cycle marker, magenta) are shown at higher magnification. 1: maxillary process, 2: mandibular arch, 3: hyoid arch, 4: heart, 5: left forelimb, 5′: right forelimb, 6: left hindlimb, 6′: right hindlimb, 7: trigeminal ganglion, 8: eye, 9: primary head vein, 10: otic vesicle, 11: facioacoustic ganglion complex, 12: dorsal aorta, 13: mesonephros, 14: mesentery, 15: neural tube, 16: dorsal root, 17: ventral horn. Bars in a: 1 mm, c, d: 200 µm.