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A quantitative comparison of micro-CT preparations in Dipteran flies

X-ray-based 3D-imaging techniques have gained fundamental significance in research areas ranging from taxonomy to bioengineering. There is demand for the characterisation of species-specific morphological adaptations, micro-CT (μCT) being the method of choice in small-scale animals. This has driven...

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Autores principales: Swart, Peter, Wicklein, Martina, Sykes, Dan, Ahmed, Farah, Krapp, Holger G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5175214/
https://www.ncbi.nlm.nih.gov/pubmed/28000717
http://dx.doi.org/10.1038/srep39380
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author Swart, Peter
Wicklein, Martina
Sykes, Dan
Ahmed, Farah
Krapp, Holger G.
author_facet Swart, Peter
Wicklein, Martina
Sykes, Dan
Ahmed, Farah
Krapp, Holger G.
author_sort Swart, Peter
collection PubMed
description X-ray-based 3D-imaging techniques have gained fundamental significance in research areas ranging from taxonomy to bioengineering. There is demand for the characterisation of species-specific morphological adaptations, micro-CT (μCT) being the method of choice in small-scale animals. This has driven the development of suitable staining techniques to improve absorption-based tissue contrast. A quantitative account on the limits of current staining protocols for preparing μCT specimen, however, is still missing. Here we present a study that quantifies results obtained by combining a variety of different contrast agents and fixative treatments that provides general guidance for μCT applications, particularly suitable for insect species. Using a blowfly model system (Calliphora), we enhanced effective spatial resolution and, in particular, optimised tissue contrast enabling semi-automated segmentation of soft and hard tissue from μCT data. We introduce a novel probabilistic measure of the contrast between tissues: PTC. Our results show that a strong iodine solution provides the greatest overall increase in tissue contrast, however phosphotungstic acid offers better inter-tissue discriminability. We further show that using paraformaldehyde as a fixative as opposed to ethanol, slows down the uptake of a staining solution by approximately a factor of two.
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spelling pubmed-51752142016-12-28 A quantitative comparison of micro-CT preparations in Dipteran flies Swart, Peter Wicklein, Martina Sykes, Dan Ahmed, Farah Krapp, Holger G. Sci Rep Article X-ray-based 3D-imaging techniques have gained fundamental significance in research areas ranging from taxonomy to bioengineering. There is demand for the characterisation of species-specific morphological adaptations, micro-CT (μCT) being the method of choice in small-scale animals. This has driven the development of suitable staining techniques to improve absorption-based tissue contrast. A quantitative account on the limits of current staining protocols for preparing μCT specimen, however, is still missing. Here we present a study that quantifies results obtained by combining a variety of different contrast agents and fixative treatments that provides general guidance for μCT applications, particularly suitable for insect species. Using a blowfly model system (Calliphora), we enhanced effective spatial resolution and, in particular, optimised tissue contrast enabling semi-automated segmentation of soft and hard tissue from μCT data. We introduce a novel probabilistic measure of the contrast between tissues: PTC. Our results show that a strong iodine solution provides the greatest overall increase in tissue contrast, however phosphotungstic acid offers better inter-tissue discriminability. We further show that using paraformaldehyde as a fixative as opposed to ethanol, slows down the uptake of a staining solution by approximately a factor of two. Nature Publishing Group 2016-12-21 /pmc/articles/PMC5175214/ /pubmed/28000717 http://dx.doi.org/10.1038/srep39380 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Swart, Peter
Wicklein, Martina
Sykes, Dan
Ahmed, Farah
Krapp, Holger G.
A quantitative comparison of micro-CT preparations in Dipteran flies
title A quantitative comparison of micro-CT preparations in Dipteran flies
title_full A quantitative comparison of micro-CT preparations in Dipteran flies
title_fullStr A quantitative comparison of micro-CT preparations in Dipteran flies
title_full_unstemmed A quantitative comparison of micro-CT preparations in Dipteran flies
title_short A quantitative comparison of micro-CT preparations in Dipteran flies
title_sort quantitative comparison of micro-ct preparations in dipteran flies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5175214/
https://www.ncbi.nlm.nih.gov/pubmed/28000717
http://dx.doi.org/10.1038/srep39380
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