{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,11]],"date-time":"2026-07-11T11:33:55Z","timestamp":1783769635453,"version":"3.55.0"},"reference-count":92,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2020,11,3]],"date-time":"2020-11-03T00:00:00Z","timestamp":1604361600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2020,11,3]],"date-time":"2020-11-03T00:00:00Z","timestamp":1604361600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001681","name":"Abdus Salam International Centre for Theoretical Physics","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001681","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Amphibians are models for studying applied ecological issues such as habitat loss, pollution, disease, and global climate change due to their sensitivity and vulnerability to changes in the environment. Developmental series of amphibians are informative about their biology, and X-ray based 3D reconstruction holds promise for quantifying morphological changes during growth\u2014some with a direct impact on the possibility of an experimental investigation on several of the ecological topics listed above. However, 3D resolution and discrimination of their soft tissues have been difficult with traditional X-ray computed tomography, without time-consuming contrast staining. Tomographic data were initially performed (pre-processing and reconstruction) using the open-source software tool SYRMEP Tomo Project. Data processing and analysis of the reconstructed tomography volumes were conducted using the segmentation semi-automatic settings of the software Avizo Fire 8, which provide information about each investigated tissues, organs or bone elements. Hence, volumetric analyses were carried out to quantify the development of structures in different tadpole developmental stages. Our work shows that synchrotron X-ray microtomography using phase-contrast mode resolves the edges of the internal tissues (as well as overall tadpole morphology), facilitating the segmentation of the investigated tissues. Reconstruction algorithms and segmentation software played an important role in the qualitative and quantitative analysis of each target structure of the<jats:italic>Thoropa miliaris<\/jats:italic>tadpole at different stages of development, providing information on volume, shape and length. The use of the synchrotron X-ray microtomography setup of the SYRMEP beamline of Elettra Synchrotron, in phase-contrast mode, allows access to volumetric data for bone formation, eye development, nervous system and notochordal changes during the development (ontogeny) of tadpoles of a cycloramphid frog<jats:italic>Thoropa miliaris<\/jats:italic>. As key elements in the normal development of these and any other frog tadpole, the application of such a comparative ontogenetic study, may hold interest to researchers in experimental and environmental disciplines.<\/jats:p>","DOI":"10.1038\/s41598-020-75993-8","type":"journal-article","created":{"date-parts":[[2020,11,3]],"date-time":"2020-11-03T11:02:41Z","timestamp":1604401361000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Synchrotron microtomography applied to the volumetric analysis of internal structures of Thoropa miliaris tadpoles"],"prefix":"10.1038","volume":"10","author":[{"given":"G.","family":"Fidalgo","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K.","family":"Paiva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G.","family":"Mendes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R.","family":"Barcellos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G.","family":"Cola\u00e7o","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G.","family":"Sena","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Pickler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C. L.","family":"Mota","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G.","family":"Tromba","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L. P.","family":"Nogueira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D.","family":"Braz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"H. R.","family":"Silva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M. V.","family":"Cola\u00e7o","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R. C.","family":"Barroso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,11,3]]},"reference":[{"issue":"6","key":"75993_CR1","doi-asserted-by":"publisher","first-page":"889","DOI":"10.1111\/joa.12449","volume":"228","author":"PM Gignac","year":"2016","unstructured":"Gignac, P. M. et al. Diffusible iodine-based contrast-enhanced computed tomography (diceCT): An emerging tool for rapid, high-resolution, 3-D imaging of metazoan soft tissues. J. Anat. 228(6), 889\u2013909 (2016).","journal-title":"J. Anat."},{"issue":"1","key":"75993_CR2","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1186\/1472-6793-9-11","volume":"9","author":"BD Metscher","year":"2009","unstructured":"Metscher, B. D. MicroCT for comparative morphology: simple staining methods allow high-contrast 3D imaging of diverse non-mineralized animal tissues. BMC Physiol. 9(1), 11 (2009).","journal-title":"BMC Physiol."},{"issue":"3","key":"75993_CR3","doi-asserted-by":"publisher","first-page":"632","DOI":"10.1002\/dvdy.21857","volume":"238","author":"BD Metscher","year":"2009","unstructured":"Metscher, B. D. MicroCT for developmental biology: A versatile tool for high-contrast 3D imaging at histological resolutions. Dev. Dyn. 238(3), 632\u2013640 (2009).","journal-title":"Dev. Dyn."},{"issue":"2-3","key":"75993_CR4","doi-asserted-by":"publisher","first-page":"104","DOI":"10.1016\/j.micron.2011.10.002","volume":"43","author":"R Mizutani","year":"2012","unstructured":"Mizutani, R. & Suzuki, Y. X-ray microtomography in biology. Micron 43(2\u20133), 104\u2013115 (2012).","journal-title":"Micron"},{"issue":"1","key":"75993_CR5","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1635\/053.165.0104","volume":"165","author":"TP Carvalho","year":"2016","unstructured":"Carvalho, T. P., Lundberg, J. G., Baskin, J. N., Friel, J. P. & Reis, R. E. A new species of the blind and miniature genus Micromyzon Friel and Lundberg, 1996 (Silurifomes: Aspredinidae) from the Orinoco River: Describing catfish diversity using high-resolution computed tomography. Proc. Acad. Nat. Sci. Phila. 165(1), 37\u201353 (2016).","journal-title":"Proc. Acad. Nat. Sci. Phila."},{"issue":"130","key":"75993_CR6","doi-asserted-by":"publisher","first-page":"20160898","DOI":"10.1098\/rsif.2016.0898","volume":"14","author":"H Herzog","year":"2017","unstructured":"Herzog, H., Klein, B. & Ziegler, A. Form and function of the teleost lateral line revealed using three-dimensional imaging and computational fluid dynamics. J. R. Soc. Interface 14(130), 20160898 (2017).","journal-title":"J. R. Soc. Interface"},{"issue":"1","key":"75993_CR7","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41598-018-34848-z","volume":"8","author":"V Weinhardt","year":"2018","unstructured":"Weinhardt, V. et al. Quantitative morphometric analysis of adult teleost fish by X-ray computed tomography. Sci. Rep. 8(1), 1\u201312 (2018).","journal-title":"Sci. Rep."},{"issue":"1","key":"75993_CR8","doi-asserted-by":"publisher","first-page":"243","DOI":"10.1111\/bij.12314","volume":"113","author":"A Ivanovic","year":"2014","unstructured":"Ivanovic, A. & Arntzen, J. W. Evolution of skull and body shape in Triturus newts reconstructed from three-dimensional morphometric data and phylogeny. Biol. J. Lin. Soc. 113(1), 243\u2013255 (2014).","journal-title":"Biol. J. Lin. Soc."},{"issue":"1","key":"75993_CR9","doi-asserted-by":"publisher","first-page":"e0211069","DOI":"10.1371\/journal.pone.0211069","volume":"14","author":"J Jia","year":"2019","unstructured":"Jia, J., Jiang, J. P., Zhang, M. H. & Gao, K. Q. Osteology of Batrachuperus yenyuanensis (Urodela, Hynobiidae), a high-altitude mountain stream salamander from western China. PLoS ONE 14(1), e0211069 (2019).","journal-title":"PLoS ONE"},{"key":"75993_CR10","doi-asserted-by":"publisher","first-page":"e4517","DOI":"10.7717\/peerj.4517","volume":"6","author":"J Jiang","year":"2018","unstructured":"Jiang, J., Jia, J., Zhang, M. & Gao, K. Q. Osteology of Batrachuperus londongensis (Urodela, Hynobiidae): Study of bony anatomy of a facultatively neotenic salamander from Mount Emei, Sichuan Province, China. PeerJ 6, e4517 (2018).","journal-title":"PeerJ"},{"issue":"12","key":"75993_CR11","doi-asserted-by":"publisher","first-page":"e50743","DOI":"10.1371\/journal.pone.0050743","volume":"7","author":"HC Maddin","year":"2012","unstructured":"Maddin, H. C., Jenkins, F. A. & Anderson, J. S. The braincase of Eocaecilia micropodia (Lissamphibia, Gymnophiona) and the origin of caecilians. PLoS ONE 7(12), e50743 (2012).","journal-title":"PLoS ONE"},{"issue":"4","key":"75993_CR12","doi-asserted-by":"publisher","first-page":"721","DOI":"10.1093\/biolinnean\/blz001","volume":"126","author":"AF Marshall","year":"2019","unstructured":"Marshall, A. F. et al. High-density three-dimensional morphometric analyses support conserved static (intraspecific) modularity in caecilian (Amphibia: Gymnophiona) crania. Biol. J. Lin. Soc. 126(4), 721\u2013742 (2019).","journal-title":"Biol. J. Lin. Soc."},{"issue":"4","key":"75993_CR13","doi-asserted-by":"publisher","first-page":"528","DOI":"10.1007\/s11692-014-9287-2","volume":"41","author":"E Sherratt","year":"2014","unstructured":"Sherratt, E., Gower, D. J., Klingenberg, C. P. & Wilkinson, M. Evolution of cranial shape in Caecilians (Amphibia: Gymnophiona). Evol. Biol. 41(4), 528\u2013545 (2014).","journal-title":"Evol. Biol."},{"key":"75993_CR14","doi-asserted-by":"publisher","first-page":"41","DOI":"10.11646\/zootaxa.2874.1.3","volume":"64","author":"M Wilkinson","year":"2011","unstructured":"Wilkinson, M., Mauro, D. S., Sherratt, E. & Gower, D. J. A nine-family classification of caecilians (Amphibia: Gymnophiona). Zootaxa 64, 41\u201364 (2011).","journal-title":"Zootaxa"},{"key":"75993_CR15","doi-asserted-by":"publisher","first-page":"e7003","DOI":"10.7717\/peerj.7003","volume":"7","author":"AJ Collings","year":"2019","unstructured":"Collings, A. J. & Richards, C. T. Digital dissection of the pelvis and hindlimb of the red-legged running frog, Phlyctimantis maculatus, using diffusible iodine contrast enhanced computed microtomography (DICE \u03bcCT). PeerJ 7, e7003 (2019).","journal-title":"PeerJ"},{"issue":"1","key":"75993_CR16","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41598-017-12145-5","volume":"7","author":"S Goutte","year":"2017","unstructured":"Goutte, S. et al. Evidence of auditory insensitivity to vocalization frequencies in two frogs. Sci. Rep. 7(1), 1\u20139 (2017).","journal-title":"Sci. Rep."},{"issue":"1","key":"75993_CR17","doi-asserted-by":"publisher","first-page":"893","DOI":"10.3762\/bjnano.7.81","volume":"7","author":"T Kleinteich","year":"2016","unstructured":"Kleinteich, T. & Gorb, S. N. Frog tongue surface microstructures: Functional and evolutionary patterns. Beilst. J. Nanotechnol. 7(1), 893\u2013903 (2016).","journal-title":"Beilst. J. Nanotechnol."},{"key":"75993_CR18","doi-asserted-by":"publisher","first-page":"e3603","DOI":"10.7717\/peerj.3603","volume":"5","author":"LF Ribeiro","year":"2017","unstructured":"Ribeiro, L. F., Blackburn, D. C., Stanley, E. L., Pie, M. R. & Bornschein, M. R. Two new species of the Brachycephalus pernix group (Anura: Brachycephalidae) from the state of Paran\u00e1, southern Brazil. PeerJ 5, e3603 (2017).","journal-title":"PeerJ"},{"key":"75993_CR19","doi-asserted-by":"publisher","first-page":"105","DOI":"10.3897\/zse.91.4979","volume":"91","author":"MD Scherz","year":"2015","unstructured":"Scherz, M. D., Rakotoarison, A., Hawlitschek, O., Vences, M. & Glaw, F. Leaping towards a saltatorial lifestyle? An unusually long-legged new species of Rhombophryne (Anura, Microhylidae) from the Sorata massif in northern Madagascar. Zoosystem. Evol. 91, 105\u2013114 (2015).","journal-title":"Zoosystem. Evol."},{"key":"75993_CR20","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1016\/j.jmbbm.2017.06.007","volume":"74","author":"C Broeckhoven","year":"2017","unstructured":"Broeckhoven, C., du Plessis, A. & Hui, C. Functional trade-off between strength and thermal capacity of dermal armor: Insights from girdled lizards. J. Mech. Behav. Biomed. Mater. 74, 189\u2013194 (2017).","journal-title":"J. Mech. Behav. Biomed. Mater."},{"issue":"137","key":"75993_CR21","doi-asserted-by":"publisher","first-page":"20170637","DOI":"10.1098\/rsif.2017.0637","volume":"14","author":"MEH Jones","year":"2017","unstructured":"Jones, M. E. H. et al. The biomechanical role of the chondrocranium and sutures in a lizard cranium. J. R. Soc. Interface 14(137), 20170637 (2017).","journal-title":"J. R. Soc. Interface"},{"issue":"10","key":"75993_CR22","doi-asserted-by":"publisher","first-page":"1675","DOI":"10.1002\/ar.24197","volume":"302","author":"JA Maisano","year":"2019","unstructured":"Maisano, J. A., Laduc, T. J., Bell, C. J. & Barber, D. The cephalic osteoderms of Varanus komodoensis as revealed by high-resolution X-ray computed tomography. Anat. Record 302(10), 1675\u20131680 (2019).","journal-title":"Anat. Record"},{"issue":"8","key":"75993_CR23","doi-asserted-by":"publisher","first-page":"20170293","DOI":"10.1098\/rsbl.2017.0293","volume":"13","author":"C Broeckhoven","year":"2017","unstructured":"Broeckhoven, C. & du Plessis, A. Has snake fang evolution lost its bite? New insights from a structural mechanics viewpoint. Biol. Lett. 13(8), 20170293 (2017).","journal-title":"Biol. Lett."},{"issue":"1","key":"75993_CR24","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41467-017-02088-w","volume":"9","author":"FO da Silva","year":"2018","unstructured":"da Silva, F. O. et al. The ecological origins of snakes as revealed by skull evolution. Nature Communications 9(1), 1\u201311 (2018).","journal-title":"Nature Communications"},{"issue":"1","key":"75993_CR25","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1093\/gigascience\/gix126","volume":"7","author":"A du Plessis","year":"2018","unstructured":"du Plessis, A., Broeckhoven, C. & le Roux, S. G. Snake fangs: 3D morphological and mechanical analysis by microCT, simulation, and physical compression testing. GigaScience 7(1), 126 (2018).","journal-title":"GigaScience"},{"issue":"1","key":"75993_CR26","doi-asserted-by":"publisher","first-page":"70","DOI":"10.1002\/jmor.10388","volume":"267","author":"JA Maisano","year":"2006","unstructured":"Maisano, J. A., Kearney, M. & Rowe, T. Cranial anatomy of the spade-headed amphisbaenian Diplometopon zarudnyi (Squamata, Amphisbaenia) based on high-resolution X-ray computed tomography. J. Morphol. 267(1), 70\u2013102 (2006).","journal-title":"J. Morphol."},{"issue":"8","key":"75993_CR27","doi-asserted-by":"publisher","first-page":"1407","DOI":"10.1002\/ar.22930","volume":"297","author":"CC Urben","year":"2014","unstructured":"Urben, C. C., Daza, J. D., Cadena, C., Lewis, P. J. & Thies, M. L. The homology of the pelvic elements of Zygaspis quadrifrons (squamata: Amphisbaenia). Anat. Record 297(8), 1407\u20131413 (2014).","journal-title":"Anat. Record"},{"issue":"1","key":"75993_CR28","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12862-018-1303-1","volume":"19","author":"N Westphal","year":"2019","unstructured":"Westphal, N., Mahlow, K., Head, J. J. & M\u00fcller, J. Pectoral myology of limb-reduced worm lizards (Squamata, Amphisbaenia) suggests decoupling of the musculoskeletal system during the evolution of body elongation. BMC Evol. Biol. 19(1), 1\u201323 (2019).","journal-title":"BMC Evol. Biol."},{"key":"75993_CR29","doi-asserted-by":"publisher","first-page":"1192","DOI":"10.1002\/jmor.10974","volume":"272","author":"G Rivera","year":"2011","unstructured":"Rivera, G. & Stayton, C. T. Finite element modeling of shell shape in the freshwater turtle Pseudemys concinna reveals a trade-off between mechanical strength and hydrodynamic efficiency. J. Morphol. 272, 1192\u20131203 (2011).","journal-title":"J. Morphol."},{"issue":"2","key":"75993_CR30","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1016\/j.rvsc.2005.12.009","volume":"81","author":"A Arencibia","year":"2006","unstructured":"Arencibia, A. et al. Computed tomographic anatomy of the head of the loggerhead sea turtle (Caretta caretta). Res. Vet. Sci. 81(2), 165\u2013169 (2006).","journal-title":"Res. Vet. Sci."},{"issue":"4","key":"75993_CR31","doi-asserted-by":"publisher","first-page":"5890","DOI":"10.1016\/j.actbio.2012.12.023","volume":"9","author":"B Achrai","year":"2013","unstructured":"Achrai, B. & Wagner, H. D. Micro-structure and mechanical properties of the turtle carapace as a biological composite shield. Acta Biomater. 9(4), 5890\u20135902 (2013).","journal-title":"Acta Biomater."},{"issue":"4","key":"75993_CR32","doi-asserted-by":"publisher","first-page":"e0175079","DOI":"10.1371\/journal.pone.0175079","volume":"12","author":"AJ Klinkhamer","year":"2017","unstructured":"Klinkhamer, A. J., Wilhite, D. R., White, M. A. & Wroe, S. Digital dissection and three-dimensional interactive models of limb musculature in the Australian estuarine crocodile (Crocodylus porosus). PLoS ONE 12(4), e0175079 (2017).","journal-title":"PLoS ONE"},{"key":"75993_CR33","doi-asserted-by":"publisher","first-page":"e988","DOI":"10.7717\/peerj.988","volume":"3","author":"MR McCurry","year":"2015","unstructured":"McCurry, M. R., Evans, A. R. & McHenry, C. R. The sensitivity of biological finite element models to the resolution of surface geometry: A case study of crocodilian crania. PeerJ 3, e988 (2015).","journal-title":"PeerJ"},{"key":"75993_CR34","doi-asserted-by":"publisher","first-page":"e60","DOI":"10.7717\/peerj.60","volume":"1","author":"ER Schachner","year":"2013","unstructured":"Schachner, E. R., Hutchinson, J. R. & Farmer, C. Pulmonary anatomy in the Nile crocodile and the evolution of unidirectional airflow in Archosauria. PeerJ 1, e60 (2013).","journal-title":"PeerJ"},{"key":"75993_CR35","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/srep16002","volume":"5","author":"D Jirak","year":"2015","unstructured":"Jirak, D., Janacek, J. & Kear, B. P. A combined MR and CT study for precise quantitative analysis of the avian brain. Sci. Rep. 5, 1\u20137 (2015).","journal-title":"Sci. Rep."},{"key":"75993_CR36","doi-asserted-by":"publisher","first-page":"273","DOI":"10.1016\/j.jmbbm.2018.05.001","volume":"84","author":"JY Jung","year":"2018","unstructured":"Jung, J. Y. et al. A comparative analysis of the avian skull: Woodpeckers and chickens. J. Mech. Behav. Biomed. Mater. 84, 273\u2013280 (2018).","journal-title":"J. Mech. Behav. Biomed. Mater."},{"key":"75993_CR37","doi-asserted-by":"publisher","first-page":"412","DOI":"10.1111\/joa.12153","volume":"224","author":"S Lautenschlager","year":"2014","unstructured":"Lautenschlager, S., Bright, J. A. & Rayfield, E. J. Digital dissection\u2014using contrast-enhanced computed tomography scanning to elucidate hard- and soft-tissue anatomy in the Common Buzzard Buteo buteo. J. Anat. 224, 412\u2013431 (2014).","journal-title":"J. Anat."},{"key":"75993_CR38","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1016\/j.aanat.2015.08.005","volume":"208","author":"X Lihong","year":"2016","unstructured":"Lihong, X. et al. Micro-computed tomography and microdissection of the temporal bone of tree shrews. Ann. Anat.-Anatomischer Anzeiger 208, 69\u201377 (2016).","journal-title":"Ann. Anat.-Anatomischer Anzeiger"},{"key":"75993_CR39","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/srep42847","volume":"7","author":"M T\u00f6pperwien","year":"2017","unstructured":"T\u00f6pperwien, M. et al. Three-dimensional mouse brain cytoarchitecture revealed by laboratory-based X-ray phase-contrast tomography. Sci. Rep. 7, 1\u20138 (2017).","journal-title":"Sci. Rep."},{"issue":"6","key":"75993_CR40","doi-asserted-by":"publisher","first-page":"859","DOI":"10.1111\/joa.12606","volume":"230","author":"B Zeller-Plumhoff","year":"2017","unstructured":"Zeller-Plumhoff, B. et al. Phase-contrast synchrotron radiation computed tomography of muscle spindles in the mouse soleus muscle. J. Anat. 230(6), 859\u2013865 (2017).","journal-title":"J. Anat."},{"issue":"6","key":"75993_CR41","doi-asserted-by":"publisher","first-page":"347","DOI":"10.1016\/j.tree.2014.04.004","volume":"29","author":"JA Cunningham","year":"2014","unstructured":"Cunningham, J. A., Rahman, I. A., Lautenschlager, S., Rayfield, E. J. & Donoghue, P. C. J. A virtual world of paleontology. Trends Ecol. Evol. 29(6), 347\u2013357 (2014).","journal-title":"Trends Ecol. Evol."},{"key":"75993_CR42","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1017\/scs.2017.6","volume":"22","author":"R Racicot","year":"2017","unstructured":"Racicot, R. Fossil secrets revealed: X-ray CT scanning and applications in paleontology. Paleontol. Soc. Pap. 22, 21\u201338 (2017).","journal-title":"Paleontol. Soc. Pap."},{"key":"75993_CR43","doi-asserted-by":"publisher","first-page":"377","DOI":"10.1163\/15685381-20181102","volume":"39","author":"C Broeckhoven","year":"2018","unstructured":"Broeckhoven, C. & du Plessis, A. X-ray microtomography in herpetological research: A review. Amphibia-Reptilia 39, 377\u2013401 (2018).","journal-title":"Amphibia-Reptilia"},{"issue":"1","key":"75993_CR44","first-page":"20","volume":"144","author":"E Descamps","year":"2014","unstructured":"Descamps, E. et al. Soft tissue discrimination with contrast agents using micro-CT scanning. Belg. J. Zool. 144(1), 20\u201340 (2014).","journal-title":"Belg. J. Zool."},{"issue":"4","key":"75993_CR45","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1089\/ten.tec.2008.0274","volume":"14","author":"R Mizutani","year":"2008","unstructured":"Mizutani, R. et al. X-ray Microtomographic imaging of three-dimensional structure of soft tissues. Tissue Eng. Part C Methods 14(4), 359\u2013363 (2008).","journal-title":"Tissue Eng. Part C Methods"},{"key":"75993_CR46","volume-title":"Tadpoles: the Biology of Anuran Larvae","author":"RW McDiarmid","year":"1999","unstructured":"McDiarmid, R. W. & Altig, R. Tadpoles: the Biology of Anuran Larvae (University of Chicago Press, Chicago, 1999)."},{"issue":"2","key":"75993_CR47","first-page":"99","volume":"9","author":"G Shu","year":"2018","unstructured":"Shu, G. et al. A rapid, non-invasive method for anatomical observations of tadpole vertebrae in vivo. Asian Herpetol. Res. 9(2), 99\u2013109 (2018).","journal-title":"Asian Herpetol. Res."},{"issue":"4","key":"75993_CR48","first-page":"22","volume":"16","author":"J Hanken","year":"1981","unstructured":"Hanken, J. & Wassersug, R. The visible skeleton. Funct. Photogr. 16(4), 22\u201326 (1981).","journal-title":"Funct. Photogr."},{"issue":"2","key":"75993_CR49","first-page":"107","volume":"9","author":"WR Taylor","year":"1985","unstructured":"Taylor, W. R. & Van Dyke, G. C. Revised procedures for staining and clearing small fishes and other vertebrates for bone and cartilage study. Cybium 9(2), 107\u2013119 (1985).","journal-title":"Cybium"},{"issue":"2","key":"75993_CR50","doi-asserted-by":"publisher","first-page":"131","DOI":"10.3109\/10520297609116684","volume":"51","author":"RJ Wassersug","year":"1976","unstructured":"Wassersug, R. J. A procedure for differential staining of cartilage and bone in whole formalin-fixed vertebrates. Stain Technol. 51(2), 131\u2013134 (1976).","journal-title":"Stain Technol."},{"key":"75993_CR51","doi-asserted-by":"publisher","first-page":"595","DOI":"10.1080\/10520295.2017.1369162","volume":"92","author":"M Krings","year":"2017","unstructured":"Krings, M., M\u00fcller, H., Heneka, M. J. & R\u00f6dder, D. Modern morphological methods for tadpole studies. A comparison of micro-CT, and clearing and staining protocols modified for frog larvae. Biotech. Histochem. 92, 595\u2013605 (2017).","journal-title":"Biotech. Histochem."},{"key":"75993_CR52","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1002\/(SICI)1097-4687(200001)243:1<75::AID-JMOR4>3.0.CO;2-L","volume":"243","author":"L Trueb","year":"2000","unstructured":"Trueb, L., P\u00fagener, L. A. & Maglia, A. M. Ontogeny of the bizarre: An osteological description of Pipa pipa (Anura: Pipidae), with an account of skeletal development in the species. J. Morphol. 243, 75\u2013104 (2000).","journal-title":"J. Morphol."},{"issue":"13","key":"75993_CR53","doi-asserted-by":"publisher","first-page":"R29","DOI":"10.1088\/0031-9155\/51\/13\/R03","volume":"51","author":"WA Kalender","year":"2006","unstructured":"Kalender, W. A. X-ray computed tomography. Phys. Med. Biol. 51(13), R29-43 (2006).","journal-title":"Phys. Med. Biol."},{"key":"75993_CR54","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1146\/annurev-bioeng-071910-124717","volume":"13","author":"EL Ritman","year":"2011","unstructured":"Ritman, E. L. Current status of developments and applications of micro-CT. Annu. Rev. Biomed. Eng. 13, 531\u2013552 (2011).","journal-title":"Annu. Rev. Biomed. Eng."},{"issue":"6","key":"75993_CR55","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/scrt534","volume":"5","author":"JD Boerckel","year":"2014","unstructured":"Boerckel, J. D., Mason, D. E., McDermott, A. M. & Alsberg, E. Microcomputed tomography: Approaches and applications in bioengineering. Stem Cell Res. Ther. 5(6), 1\u201312 (2014).","journal-title":"Stem Cell Res. Ther."},{"issue":"6515","key":"75993_CR56","doi-asserted-by":"publisher","first-page":"595","DOI":"10.1038\/373595a0","volume":"373","author":"TJ Davis","year":"1995","unstructured":"Davis, T. J., Gao, D., Gureyev, T. E., Stevenson, A. W. & Wilkins, S. W. Phase-contrast imaging of weakly absorbing materials using hard X-rays. Nature 373(6515), 595\u2013598 (1995).","journal-title":"Nature"},{"issue":"6607","key":"75993_CR57","doi-asserted-by":"publisher","first-page":"335","DOI":"10.1038\/384335a0","volume":"384","author":"SW Wilkins","year":"1996","unstructured":"Wilkins, S. W., Gureyev, T. E., Gao, D., Pogany, A. & Stevenson, A. W. Phase-contrast imaging using polychromatic hard X-rays. Nature 384(6607), 335\u2013338 (1996).","journal-title":"Nature"},{"issue":"7","key":"75993_CR58","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1063\/1.1292471","volume":"53","author":"R Fitzgerald","year":"2000","unstructured":"Fitzgerald, R. Phase-sensitive X-ray imaging. Phys. Today 53(7), 23\u201326 (2000).","journal-title":"Phys. Today"},{"issue":"9R","key":"75993_CR59","doi-asserted-by":"publisher","first-page":"6355","DOI":"10.1143\/JJAP.44.6355","volume":"44","author":"A Momose","year":"2005","unstructured":"Momose, A. Recent advances in X-ray phase imaging. Jpn. J. Appl. Phys. 44(9R), 6355\u20136367 (2005).","journal-title":"Jpn. J. Appl. Phys."},{"issue":"4","key":"75993_CR60","doi-asserted-by":"publisher","first-page":"258","DOI":"10.1038\/nphys265","volume":"2","author":"F Pfeiffer","year":"2006","unstructured":"Pfeiffer, F., Weitkamp, T., Bunk, O. & David, C. Phase retrieval and differential phase-contrast imaging with low-brilliance X-ray sources. Nat. Phys. 2(4), 258\u2013261 (2006).","journal-title":"Nat. Phys."},{"issue":"5","key":"75993_CR61","doi-asserted-by":"publisher","first-page":"3223","DOI":"10.1364\/OE.16.003223","volume":"16","author":"TE Gureyev","year":"2008","unstructured":"Gureyev, T. E. et al. Some simple rules for contrast, signal-to-noise and resolution in in-line X-ray phase-contrast imaging. Opt. Express 16(5), 3223\u20133241 (2008).","journal-title":"Opt. Express"},{"key":"75993_CR62","doi-asserted-by":"publisher","first-page":"20130021","DOI":"10.1098\/rsta.2013.0021","volume":"372","author":"SW Wilkins","year":"2014","unstructured":"Wilkins, S. W. et al. On the evolution and relative merits of hard X-ray phase-contrast imaging methods. Philos. Trans. R. Soc. A Math. Phys. Eng. Sci. 372, 20130021 (2014).","journal-title":"Philos. Trans. R. Soc. A Math. Phys. Eng. Sci."},{"key":"75993_CR63","doi-asserted-by":"publisher","first-page":"88","DOI":"10.1016\/j.nima.2017.07.036","volume":"878","author":"M Endrizzi","year":"2018","unstructured":"Endrizzi, M. X-ray phase-contrast imaging. Nucl. Instrum. Methods Phys. Res., Sect. A 878, 88\u201398 (2018).","journal-title":"Nucl. Instrum. Methods Phys. Res., Sect. A"},{"issue":"13","key":"75993_CR64","doi-asserted-by":"publisher","first-page":"R105","DOI":"10.1088\/0022-3727\/35\/13\/201","volume":"35","author":"Y Hwu","year":"2002","unstructured":"Hwu, Y., Tsai, W. L., Groso, A., Margaritondo, G. & Je, J. H. Coherence-enhanced synchrotron radiology: Simple theory and practical applications. J. Phys. D Appl. Phys. 35(13), R105\u2013R120 (2002).","journal-title":"J. Phys. D Appl. Phys."},{"issue":"2-3","key":"75993_CR65","doi-asserted-by":"publisher","first-page":"280","DOI":"10.1504\/IJNT.2006.009584","volume":"3","author":"BM Weon","year":"2006","unstructured":"Weon, B. M., Je, J. H., Hwu, Y. & Margaritondo, G. Phase contrast X-ray imaging. Int. J. Nanotechnol. 3(2\u20133), 280\u2013297 (2006).","journal-title":"Int. J. Nanotechnol."},{"issue":"1","key":"75993_CR66","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1088\/0022-3727\/29\/1\/023","volume":"29","author":"P Cloetens","year":"1996","unstructured":"Cloetens, P., Barrett, R., Baruchel, J., Guigay, J. P. & Schlenker, M. Phase objects in synchrotron radiation hard x-ray imaging. J. Phys. D Appl. Phys. 29(1), 133\u2013146 (1996).","journal-title":"J. Phys. D Appl. Phys."},{"issue":"6","key":"75993_CR67","doi-asserted-by":"publisher","first-page":"2315","DOI":"10.1088\/1361-6560\/aa5d3d","volume":"62","author":"P Baran","year":"2017","unstructured":"Baran, P. et al. Optimization of propagation-based X-ray phase-contrast tomography for breast cancer imaging. Phys. Med. Biol. 62(6), 2315\u20132332 (2017).","journal-title":"Phys. Med. Biol."},{"key":"75993_CR68","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1111\/j.1365-2818.2007.01785.x","volume":"227","author":"O Betz","year":"2007","unstructured":"Betz, O. et al. Imaging applications of synchrotron X-ray phase-contrast microtomography in biological morphology and biomaterials science. I. General aspects of the technique and its advantages in the analysis of millimetre-sized arthropod structure. J. Microsc. 227, 51\u201371 (2007).","journal-title":"J. Microsc."},{"key":"75993_CR69","unstructured":"Brazilian National Council for Animal Experimentation Control (CONCEA). CONCEA eutan\u00e1sia practice guidelines. Brazilian Ministry of Science, Technology and Innovation (2013)."},{"key":"75993_CR70","first-page":"183","volume":"16","author":"K Gosner","year":"1960","unstructured":"Gosner, K. A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica 16, 183\u2013190 (1960).","journal-title":"Herpetologica"},{"issue":"05","key":"75993_CR71","doi-asserted-by":"publisher","first-page":"C05012","DOI":"10.1088\/1748-0221\/13\/05\/C05012","volume":"13","author":"G Fidalgo","year":"2018","unstructured":"Fidalgo, G. et al. Virtual dissection of Thoropa miliaris tadpole using phase-contrast synchrotron microtomography. J. Instrum. 13(05), C05012 (2018).","journal-title":"J. Instrum."},{"issue":"1","key":"75993_CR72","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1063\/1.3478190","volume":"1266","author":"G Tromba","year":"2010","unstructured":"Tromba, G. et al. The SYRMEP beamline of elettra: Clinical mammography and bio-medical applications. AIP Conf. Proc. 1266(1), 18\u201323 (2010).","journal-title":"AIP Conf. Proc."},{"key":"75993_CR73","doi-asserted-by":"publisher","first-page":"316","DOI":"10.1016\/j.lithos.2019.05.010","volume":"340","author":"A Liedl","year":"2019","unstructured":"Liedl, A. et al. A 3D imaging textural characterization of pyroclastic products from the 1538 AD Monte Nuovo eruption (Campi Flegrei, Italy). Lithos 340, 316\u2013331 (2019).","journal-title":"Lithos"},{"issue":"2","key":"75993_CR74","doi-asserted-by":"publisher","first-page":"81","DOI":"10.3390\/coatings9020081","volume":"9","author":"G Fiocco","year":"2019","unstructured":"Fiocco, G. et al. A micro-tomographic insight into the coating systems of historical bowed string instruments. Coatings 9(2), 81 (2019).","journal-title":"Coatings"},{"issue":"1","key":"75993_CR75","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41598-018-37186-2","volume":"9","author":"CB Cannata","year":"2019","unstructured":"Cannata, C. B. et al. First 3D imaging characterization of Pele\u2019s hair from Kilauea volcano (Hawaii). Sci. Rep. 9(1), 1\u201313 (2019).","journal-title":"Sci. Rep."},{"issue":"12","key":"75993_CR76","doi-asserted-by":"publisher","first-page":"2642","DOI":"10.1109\/TMI.2018.2845905","volume":"37","author":"P Baran","year":"2018","unstructured":"Baran, P. et al. High-resolution X-ray phase-contrast 3D imaging of breast tissue specimens as a possible adjunct to histopathology. IEEE Trans. Med. Imaging 37(12), 2642\u20132650 (2018).","journal-title":"IEEE Trans. Med. Imaging"},{"key":"75993_CR77","volume-title":"Syrmep Tomo Project tutorial","author":"F Montanari","year":"2003","unstructured":"Montanari, F. Syrmep Tomo Project tutorial (Internet report, Sincrotrone Trieste, 2003)."},{"issue":"1","key":"75993_CR78","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s40679-016-0036-8","volume":"3","author":"F Brun","year":"2017","unstructured":"Brun, F. et al. SYRMEP Tomo Project: a graphical user interface for customizing CT reconstruction workflows. Adv. Struct. Chem. Imaging 3(1), 1\u20139 (2017).","journal-title":"Adv. Struct. Chem. Imaging"},{"issue":"1","key":"75993_CR79","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1046\/j.1365-2818.2002.01010.x","volume":"206","author":"DM Paganin","year":"2002","unstructured":"Paganin, D. M. et al. Simultaneous phase and amplitude extraction from a single defocused image of a homogeneous object. J. Microsc. 206(1), 33\u201340 (2002).","journal-title":"J. Microsc."},{"key":"75993_CR80","unstructured":"Avizo. Avizo User\u2019s Guide (2016)."},{"issue":"1","key":"75993_CR81","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1186\/s13227-015-0025-3","volume":"6","author":"G Annona","year":"2015","unstructured":"Annona, G., Holland, N. D. & D\u2019Aniello, S. Evolution of the notochord. EvoDevo 6(1), 30\u201342 (2015).","journal-title":"EvoDevo"},{"issue":"16","key":"75993_CR82","doi-asserted-by":"publisher","first-page":"2989","DOI":"10.1007\/s00018-015-1897-z","volume":"72","author":"D Corallo","year":"2015","unstructured":"Corallo, D., Trapani, V. & Bonaldo, P. The notochord: Structure and functions. Cell. Mol. Life Sci. 72(16), 2989\u20133008 (2015).","journal-title":"Cell. Mol. Life Sci."},{"issue":"4","key":"75993_CR83","doi-asserted-by":"publisher","first-page":"873","DOI":"10.1242\/dev.00952","volume":"131","author":"A Fleming","year":"2004","unstructured":"Fleming, A., Keynes, R. & Tannahill, D. A central role for the notochord in vertebral patterning. Development 131(4), 873\u2013880 (2004).","journal-title":"Development"},{"issue":"4-5","key":"75993_CR84","doi-asserted-by":"publisher","first-page":"263","DOI":"10.1007\/s00429-003-0349-y","volume":"207","author":"S Grotmol","year":"2003","unstructured":"Grotmol, S., Kryvi, H., Nordvik, K. & Totland, G. K. Notochord segmentation may lay down the pathway for the development of the vertebral bodies in the Atlantic salmon. Anat. Embryol. 207(4\u20135), 263\u2013272 (2003).","journal-title":"Anat. Embryol."},{"issue":"5","key":"75993_CR85","doi-asserted-by":"publisher","first-page":"667","DOI":"10.1083\/jcb.201212095","volume":"200","author":"K Ellis","year":"2013","unstructured":"Ellis, K., Bagwell, J. & Bagnat, M. Notochord vacuoles are lysosome-related organelles that function in axis and spine morphogenesis. J. Cell Biol. 200(5), 667\u2013679 (2013).","journal-title":"J. Cell Biol."},{"key":"75993_CR86","doi-asserted-by":"publisher","first-page":"2503","DOI":"10.1242\/dev.01812","volume":"132","author":"DL Stemple","year":"2005","unstructured":"Stemple, D. L. Structure and function of the notochord: an essential organ for chordate development. Development 132, 2503\u20132512 (2005).","journal-title":"Development"},{"issue":"81","key":"75993_CR87","first-page":"171","volume":"20","author":"K Shiota","year":"1929","unstructured":"Shiota, K. Advances in the study of fetal development: From descriptive to dynamic embryology. Contribut. Embriol. 20(81), 171\u2013178 (1929).","journal-title":"Contribut. Embriol."},{"key":"75993_CR88","doi-asserted-by":"crossref","DOI":"10.56021\/9780801847806","volume-title":"Biology of amphibians","author":"WE Duellman","year":"1994","unstructured":"Duellman, W. E. & Trueb, L. Biology of amphibians (JHU press, Baltimore, 1994)."},{"key":"75993_CR89","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1007\/BF00605022","volume":"150","author":"JG Sivak","year":"1983","unstructured":"Sivak, J. G. & Warburg, M. R. Changes in optical properties of the eye during metamorphosis of an anuran, Pelobates syriacus. J. Comp. Physiol. 150, 329\u2013332 (1983).","journal-title":"J. Comp. Physiol."},{"key":"75993_CR90","doi-asserted-by":"publisher","first-page":"138","DOI":"10.1002\/jemt.22320","volume":"77","author":"AB Benjamin","year":"2014","unstructured":"Benjamin, A. B., Ahenkorah, J., Hottor, B. A., Dennis, E. & Frederick, K. A. Improved method of producing satisfactory sections of whole eyeball by routine histology. Microsc. Res. Tech. 77, 138\u2013142 (2014).","journal-title":"Microsc. Res. Tech."},{"issue":"4","key":"75993_CR91","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1136\/bjo.24.4.153","volume":"24","author":"A Gl\u00fccksmann","year":"1940","unstructured":"Gl\u00fccksmann, A. Development and differentiation of the tadpole eye. Br. J. Ophthalmol. 24(4), 153\u2013178 (1940).","journal-title":"Br. J. Ophthalmol."},{"key":"75993_CR92","doi-asserted-by":"publisher","first-page":"970","DOI":"10.1002\/neu.480210704","volume":"21","author":"SG Hoskins","year":"1990","unstructured":"Hoskins, S. G. Metamorphosis of the amphibian eye. J. Neurobiol. 21, 970\u2013989 (1990).","journal-title":"J. Neurobiol."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-75993-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-75993-8","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-75993-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,11]],"date-time":"2023-10-11T08:26:44Z","timestamp":1697012804000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-75993-8"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,3]]},"references-count":92,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2020,12]]}},"alternative-id":["75993"],"URL":"https:\/\/doi.org\/10.1038\/s41598-020-75993-8","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,11,3]]},"assertion":[{"value":"5 March 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 October 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"3 November 2020","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"18934"}}