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      Micro-CT protocols for scanning and 3D analysis of Hexaplextrunculus during its different life stages

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          Abstract

          Micro-computed tomography (micro-CT) is a high-resolution 3D-imaging technique which is now increasingly applied in biological studies focusing on taxonomy and functional morphology. The creation of virtual representations of specimens can increase availability of otherwise underexploited and inaccessible samples. The 3D model dataset can be also further processed through volume rendering and morphometric analysis. The success of micro-CT as a visualisation technique depends on several methodological manipulations, including the use of contrast enhancing staining agents, filters, scanning mediums, containers, exposure time and frame averaging. The aim of this study was to standardise a series of micro-CT scanning and 3D analysis protocols for a marine gastropod species, Hexaplex trunculus . The analytical protocols have followed all the developmental stages of this gastropod, from egg capsules and embryos to juveniles and adults.

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          Most cited references16

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          A new method for the model-independent assessment of thickness in three-dimensional images

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            MicroCT for developmental biology: a versatile tool for high-contrast 3D imaging at histological resolutions.

            Understanding developmental processes requires accurate visualization and parameterization of three-dimensional embryos. Tomographic imaging methods offer automatically aligned and calibrated volumetric images, but the usefulness of X-ray CT imaging for developmental biology has been limited by the low inherent contrast of embryonic tissues. Here, I demonstrate simple staining methods that allow high-contrast imaging of embryonic tissues at histological resolutions using a commercial microCT system. Quantitative comparisons of images of chick embryos treated with different contrast agents show that three very simple methods using inorganic iodine and phosphotungstic acid produce overall contrast and differential tissue contrast for X-ray imaging at least as high as that obtained with osmium. The stains can be used after any common fixation and after storage in aqueous or alcoholic media, and on a wide variety of species. These methods establish microCT imaging as a useful tool for comparative developmental studies, embryo phenotyping, and quantitative modeling of development. (c) 2009 Wiley-Liss, Inc.
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              Micro-computed tomography: Introducing new dimensions to taxonomy

              Abstract Continuous improvements in the resolution of three-dimensional imaging have led to an increased application of these techniques in conventional taxonomic research in recent years. Coupled with an ever increasing research effort in cybertaxonomy, three-dimensional imaging could give a boost to the development of virtual specimen collections, allowing rapid and simultaneous access to accurate virtual representations of type material. This paper explores the potential of micro-computed tomography (X-ray micro-tomography), a non-destructive three-dimensional imaging technique based on mapping X-ray attenuation in the scanned object, for supporting research in systematics and taxonomy. The subsequent use of these data as virtual type material, so-called “cybertypes”, and the creation of virtual collections lie at the core of this potential. Sample preparation, image acquisition, data processing and presentation of results are demonstrated using polychaetes (bristle worms), a representative taxon of macro-invertebrates, as a study object. Effects of the technique on the morphological, anatomical and molecular identity of the specimens are investigated. The paper evaluates the results and discusses the potential and the limitations of the technique for creating cybertypes. It also discusses the challenges that the community might face to establish virtual collections. Potential future applications of three-dimensional information in taxonomic research are outlined, including an outlook to new ways of producing, disseminating and publishing taxonomic information.
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                Author and article information

                Contributors
                Journal
                Biodivers Data J
                Biodivers Data J
                1
                urn:lsid:arphahub.com:pub:F9B2E808-C883-5F47-B276-6D62129E4FF4
                urn:lsid:zoobank.org:pub:245B00E9-BFE5-4B4F-B76E-15C30BA74C02
                Biodiversity Data Journal
                Pensoft Publishers
                1314-2836
                1314-2828
                2021
                15 September 2021
                : 9
                : e71542
                Affiliations
                [1 ] Hellenic Centre for Marine Research (HCMR), Institute of Marine Biology, Biotechnology and Aquaculture (IMBBC), Heraklion, Crete, Greece Hellenic Centre for Marine Research (HCMR), Institute of Marine Biology, Biotechnology and Aquaculture (IMBBC) Heraklion, Crete Greece
                [2 ] Biology Department, University of Crete, Heraklion, Crete, Greece Biology Department, University of Crete Heraklion, Crete Greece
                Author notes
                Corresponding author: Eva Chatzinikolaou ( evachatz@ 123456hcmr.gr ).

                Academic editor: Alexander M. Weigand

                Author information
                https://orcid.org/0000-0002-7171-5105
                https://orcid.org/0000-0002-6693-2033
                Article
                71542 17468
                10.3897/BDJ.9.e71542
                8458271
                34616212
                9be69bb9-9a66-4c11-82da-cce0a86f2997
                Eva Chatzinikolaou, Kleoniki Keklikoglou

                This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 13 July 2021
                : 06 September 2021
                Page count
                Figures: 6, Tables: 0, References: 16
                Funding
                This work was funded under the project ECCO (HFRI, Hellenic Foundation for Research and Innovation for the support of Post-doctoral Researchers, project ID 343).
                Categories
                Methods

                micro-ct,gastropod,shell,statolith,morphology,methodology,scanning, hexaplex trunculus

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