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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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Correlative 2D and 3D images of a stag beetle larva. A whole stag beetle larva was cut into sections at a thickness of 20 µm. MPR and volume rendering were reconstructed from 1064 block-face images. (a) The sagittal plane was reconstructed by MPR. (b) Sections were stained with H&E, and correlated with block-face images. The gap between the newly developing pupal cuticle and the old larval cuticle was maintained in both the block-face image and the section (asterisk). (c) Two pairs of wings are shown in the block-face image. Volume rendering shows the surface structures of the thorax, and a forewing and hindwing within the thorax. 1: fat body, 2: brain, 3: oral cavity, 4: midgut, 5: hindgut, 6: larval cuticle, 7: pupal cuticle, 8: muscle, 9: larval exocuticle, 10: larval endocuticle, 11: hair, 12: epidermis, 13: forewing, 14: hindwing, 15: mesothorax, 16: metathorax, 17: prothorax. Bars in a: 5 mm, b, c: 1 mm.