https://phaidra.vetmeduni.ac.at/o:5206 Microscopic dual-energy computed tomography (microDECT) imaging of animal tissues: the colour of X-rays Handschuh, Stephan Stephan Handschuh 0000-0002-2140-7892 University of Veterinary Medicine Vienna Reichart, Ursula Ursula Reichart 0000-0001-9709-0919 University of Veterinary Medicine Vienna Kummer, Stefan Stefan Kummer 0000-0002-0746-1403 University of Veterinary Medicine Vienna Gabner, Simone Simone Gabner 0000-0001-8853-5939 University of Veterinary Medicine Vienna Ruthensteiner, Bernhard Bernhard Ruthensteiner 0000-0002-2205-7943 Sombke, Andy Andy Sombke 0000-0001-7383-440X Schwaha, Thomas Thomas Schwaha 0000-0003-0526-6791 Beisser, Christian J. Christian J. Beisser 0009-0004-6503-5754 Lemell, Patrick Patrick Lemell 0000-0003-0262-4047 Haberthür, David David Haberthür 0000-0003-3388-9187 Hlushchuk, Ruslan Ruslan Hlushchuk 0000-0002-6722-8996 Glueckert, Rudolf Rudolf Glueckert 0000-0002-2900-6866 Metscher, Brian D. Brian D. Metscher 0000-0002-6514-4406 Glösmann, Martin Martin Glösmann 0000-0002-2094-3247 University of Veterinary Medicine Vienna Walter de Gruyter journal article open access eng In this view article, we present potential applications of laboratory-based spectral dual-energy X-ray microtomography imaging for ex vivo animal tissue samples. Technical details on this topic have been reported in previous work, so we focus explicitly on applications here, discussing both dual-energy imaging for the separation of mineralised tissue from one X-ray dense contrast agent, as well as the imaging of samples stained with two X-ray dense contrast agents. Examples are given for a variety of research areas, including preclinical biomedical phenotyping (vasculature, skeletal development), comparative vertebrate morphology, and invertebrate anatomy. Based on the data presented, we conclude that the use of spectral X-ray information can provide new insights into CT datasets. Although using the dual-energy approach initially requires more effort and cost in data acquisition, this additional effort is often worthwhile, as dual-energy datasets allow for more efficient data visualisation, segmentation, and analysis. Until now, software functions for acquisition and processing of dual-energy data have not been implemented in many of the software toolboxes provided by X-ray microtomography vendors with their scanners. We propose that a user-friendly implementation of software tools for acquiring and processing spectral CT data would be a big step towards a wider use of this promising and powerful imaging approach. X-ray Microtomography MicroCT µCT DECT Basis Material Decomposition DiceCT http://creativecommons.org/licenses/by/4.0/ https://phaidra.vetmeduni.ac.at/api/object/o:5206/download 10.1515/mim-2025-0006 https://phaidra.vetmeduni.ac.at/o:605 application/pdf Methods in Microscopy issn:2942-3899 VoR Methods in Microscopy 2 3 7.14 MB 2025