An exploratory study of contrast agents for soft tissue visualization by means of high resolution X‐ray computed tomography imaging
Abstract
Summary: High resolution X‐ray computed tomography (CT), or microCT, is a promising and already widely used technique in various scientific fields. Also for histological purposes it has great potential. Although microCT has proven to be a valuable technique for the imaging of bone structures, the visualization of soft tissue structures is still an important challenge due to their low inherent X‐ray contrast. One way to achieve contrast enhancement is to make use of contrast agents. However, contrary to light and electron microscopy, knowledge about contrast agents and staining procedures is limited for X‐ray CT. The purpose of this paper is to identify useful X‐ray contrast agents for soft tissue visualization, which can be applied in a simple way and are also suited for samples larger than (1 cm) 3 . And 28 chemical substances have been investigated. All chemicals were applied in the form of concentrated aqueous solutions in which the samples were immersed. First, strips of green Bacon were stained to evaluate contrast enhancement between muscle and adipose tissue. Furthermore it was also tested whether the contrast agents remained fixed in the tissue after staining by re‐immersing them in water. Based on the results, 12 contrast agents were selected for further testing on postmortem mice hind legs, containing a variety of different tissues, including muscle, fat, bone, cartilage and tendons. It was evaluated whether the contrast agents allowed a clearer distinction between the different soft tissue structures present. Finally also penetration depth was measured. And 26 chemicals resulted in contrast enhancement between muscle and adipose tissue in the Bacon strips. Mercury(II)chloride (HgCl 2 ), phosphotungstic acid (PTA), phosphomolybdic acid (PMA) and ammonium orthomolybdate ((NH 4 ) 2 MoO 4 ) remained fixed after re‐immersion in water. The penetration tests showed that potassium iodide (KI) and sodium tungstate can be most efficiently used for large samples of the order of several tens of cm 3 . PMA, PTA, HgCl 2 and also to a lesser extent Na 2 WO 4 and (NH 4 ) 2 MoO 4 allowed a clearer distinction between the different soft tissue structures present.
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To image insects with conventional micro-CT, staining and/or drying is often required to discriminate between tissues.
“to image insects with conventional micro-CT, staining and/or drying is often required to discriminate between tissues 16 – 19”
From High-throughput phenomics of global ant biodiversity (2026) · cites this paper as [19]
Cites
Metscher, B. D. (2009) · BMC Physiology · doi:10.1186/1472-6793-9-11 · Cited by 1,086 · see citing works
Metscher, B. (2009) · Developmental Dynamics · doi:10.1002/dvdy.21857 · Cited by 609 · see citing works
Studies on the character and staining of fibrin
Lendrum, A. C., Fraser, D., Slidders, W., & Henderson, R. (1962) · Journal of Clinical Pathology · doi:10.1136/jcp.15.5.401 · Cited by 432 · see citing works
Software tools for quantification of X-ray microtomography at the UGCT
Vlassenbroeck, J., Dierick, M., Masschaele, B., et al. (2007) · Nuclear Instruments and Methods in Physics Research Section A Accelerators Spectrometers Detectors and Associated Equipment · doi:10.1016/j.nima.2007.05.073 · Cited by 383 · see citing works
Rapid 3D Phenotyping of Cardiovascular Development in Mouse Embryos by Micro-CT With Iodine Staining
Degenhardt, K. R., Wright, A. C., Horng, D. E., et al. (2010) · Circulation Cardiovascular Imaging · doi:10.1161/circimaging.109.918482 · Cited by 267 · see citing works
A Stable Lead by Modification of Sato's Method
Hanaichi, T., Sato, T., IWAMOTO, T., Malavasi, J. Y., Hoshino, M., & Mizuno, N. (1986) · Journal of Electron Microscopy · doi:10.1093/oxfordjournals.jmicro.a050582 · Cited by 256 · see citing works
Micro-computed tomography with iodine staining resolves the arrangement of muscle fibres
Jeffery, N. S., Stephenson, R. S., Gallagher, J. A., Jarvis, J. C., & Cox, P. G. (2010) · Journal of Biomechanics · doi:10.1016/j.jbiomech.2010.08.027 · Cited by 222 · see citing works
X-ray microtomography in biology
Mizutani, R., & Suzuki, Y. (2011) · Micron · doi:10.1016/j.micron.2011.10.002 · Cited by 218 · see citing works
Histology by magnetic resonance microscopy.
Johnson, G. A., Benveniste, H., Black, R. D., Hedlund, L. W., Maronpot, R. R., & Smith, B. R. (1993) · PubMed · Cited by 214 · see citing works
UGCT: New X-ray radiography and tomography facility
Masschaele, B., Cnudde, V., Dierick, M., Jacobs, P., Hoorebeke, L. V., & Vlassenbroeck, J. (2007) · Nuclear Instruments and Methods in Physics Research Section A Accelerators Spectrometers Detectors and Associated Equipment · doi:10.1016/j.nima.2007.05.099 · Cited by 204 · see citing works
Negative staining and cryo-negative staining of macromolecules and viruses for TEM
Carlo, S. D., & Harris, J. R. (2010) · Micron · doi:10.1016/j.micron.2010.06.003 · Cited by 192 · see citing works
Thomsen, J. S., Laib, A., Koller, B. J., Prohaska, S., Mosekilde, L., & Gowin, W. (2005) · Journal of Microscopy · doi:10.1111/j.1365-2818.2005.01469.x · Cited by 188 · see citing works
Virtual Histology of Transgenic Mouse Embryos for High-Throughput Phenotyping
Johnson, J. T., Hansen, M. S., Wu, I. Q., Healy, L., Johnson, C. R., Jones, G., Capecchi, M. R., & Keller, C. (2006) · PLoS Genetics · doi:10.1371/journal.pgen.0020061 · Cited by 183 · see citing works
Cox, P. G., & Jeffery, N. S. (2011) · The Anatomical Record · doi:10.1002/ar.21381 · Cited by 167 · see citing works
Magnetic resonance histology for morphologic phenotyping
Johnson, G. A., Cofer, G. P., Fubara, B. N., Gewalt, S. L., Hedlund, L. W., & Maronpot, R. R. (2002) · Journal of Magnetic Resonance Imaging · doi:10.1002/jmri.10175 · Cited by 122 · see citing works
Magnetic resonance microscopy: recent advances and applications
Tyszka, J. M., Fraser, S. E., & Jacobs, R. E. (2004) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2004.11.004 · Cited by 122 · see citing works
Small Animal Imaging with Magnetic Resonance Microscopy
Driehuys, B., Nouls, J. C., Badea, A., Bucholz, E., Ghaghada, K. B., Petiet, A., & Hedlund, L. W. (2008) · ILAR Journal · doi:10.1093/ilar.49.1.35 · Cited by 106 · see citing works
3D micro-CT imaging of the postmortem brain
de Crespigny, A. J., Bou-Reslan, H., Nishimura, M. C., PHILLIPS, H. S., Carano, R. A. D., & D'Arceuil, H. E. (2008) · Journal of Neuroscience Methods · doi:10.1016/j.jneumeth.2008.03.006 · Cited by 103 · see citing works
Imaging honey bee brain anatomy with micro-X-ray-computed tomography
Ribi, W. A., Senden, T. J., Sakellariou, A., Limaye, A. C., & Zhang, S. (2008) · Journal of Neuroscience Methods · doi:10.1016/j.jneumeth.2008.02.010 · Cited by 90 · see citing works
Particelli, F., Mecozzi, L., Beraudi, A., Montesi, M., Baruffaldi, F., & Viceconti, M. (2011) · Journal of Microscopy · doi:10.1111/j.1365-2818.2011.03573.x · Cited by 77 · see citing works
X-ray Microcomputer Tomography for the Study of Biomineralized Endo- and Exoskeletons of Animals
Neues, F., & Epple, M. (2008) · Chemical Reviews · doi:10.1021/cr078250m · Cited by 77 · see citing works
X-Ray Microtomographic Imaging of Intact Vertebrate EmbryosPreprint
Metscher, B. (2011) · Cold Spring Harbor Protocols · doi:10.1101/pdb.prot067033 · Cited by 69 · see citing works
Watz, H., Breithecker, A., Rau, W. S., et al. (2005) · Radiology · doi:10.1148/radiol.2363041142 · Cited by 62 · see citing works
Britz, H. M., Jokihaara, J., Leppänen, O., et al. (2010) · Journal of Microscopy · doi:10.1111/j.1365-2818.2010.03381.x · Cited by 53 · see citing works
Golding, R. E., Ponder, W. F., & Byrne, M. (2008) · Journal of Morphology · doi:10.1002/jmor.10699 · Cited by 53 · see citing works
Computed tomography imaging of the neuronal structure ofDrosophilabrain
Mizutani, R., Takeuchi, A., Hara, T., Uesugi, K., & Suzuki, Y. (2007) · Journal of Synchrotron Radiation · doi:10.1107/s0909049507009004 · Cited by 52 · see citing works
Micro-Computed Tomographical Imaging of Soft Biological Materials Using Contrast Techniques
Faraj, K. A., Cuijpers, V. M. J. I., Wismans, R. G., et al. (2009) · Tissue Engineering Part C Methods · doi:10.1089/ten.tec.2008.0436 · Cited by 48 · see citing works
Virtual histology by means of high‐resolution X‐ray CT
Cnudde, V., Masschaele, B., Cock, H. E. V. D., et al. (2008) · Journal of Microscopy · doi:10.1111/j.1365-2818.2008.02142.x · Cited by 48 · see citing works
DINLEY, J., Hawkins, L. A., PATERSON, G. L. J., et al. (2009) · Journal of Microscopy · doi:10.1111/j.1365-2818.2009.03335.x · Cited by 42 · see citing works
Frisullo, P., Marino, R., Laverse, J., et al. (2010) · Meat Science · doi:10.1016/j.meatsci.2010.01.008 · Cited by 41 · see citing works
Micro-CT as a novel technique for 3D reconstruction of molluscan anatomy
Golding, R. E., & Jones, A. S. (2007) · Molluscan Research · doi:10.11646/mr.27.3.2 · Cited by 41 · see citing works
A LabVIEW® based generic CT scanner control software platform
Dierick, M., Loo, D. V., Masschaele, B., et al. (2010) · Journal of X-Ray Science and Technology · doi:10.3233/xst-2010-0268 · Cited by 40 · see citing works
Harland, D. P., Vernon, J. A., Walls, R. J., et al. (2011) · Journal of Microscopy · doi:10.1111/j.1365-2818.2011.03493.x · Cited by 32 · see citing works
Zysk, A. M., Garson, A. B., Xu, Q., et al. (2012) · Biomedical Optics Express · doi:10.1364/boe.3.001924 · Cited by 30 · see citing works
A new preparation method to study fresh plant structures with X‐ray computed tomography
Leroux, O., Leroux, F. R., Bellefroid, E., et al. (2009) · Journal of Microscopy · doi:10.1111/j.1365-2818.2008.03088.x · Cited by 28 · see citing works
Alternative staining methods for Lowicryl sections.
Horowitz, R. A., & Woodcock, C. L. F. (1992) · Journal of Histochemistry & Cytochemistry · doi:10.1177/40.1.1370308 · Cited by 26 · see citing works
A LabVIEW® based generic CT scanner control software platform
Dierick, M., Loo, D. V., Masschaele, B., et al. (2010) · Journal of X-Ray Science and Technology · doi:10.3233/xst-2010-026800268 · Cited by 6 · see citing works
Improved Methods for Staining Fibrin
Churukian, C. J. (2003) · Journal of Histotechnology · doi:10.1179/his.2003.26.2.127 · Cited by 4 · see citing works
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HANDSCHUH, S., BEISSER, C. J., RUTHENSTEINER, B., & METSCHER, B. D. (2017) · Journal of Microscopy · doi:10.1111/jmi.12543 · Cited by 48 · see citing works
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Katzke, J., Hita Garcia, F., Lösel, P. D., Azuma, F., Faragó, T., Aibekova, L., Casadei-Ferreira, A., Gautam, S., Richter, A., Toulkeridou, E., et al. (2026) · Nature Methods · doi:10.1038/s41592-026-03005-0 · Cited by 6 · see citing works
“However, to image insects with conventional micro-CT, staining and/or drying is often required to discriminate between tissues 16–19 , which not only necessitates additional preprocessing steps but alters specimens and excludes some collection material from imaging.”
- Authors
- PAUWELS, E., VAN LOO, D., CORNILLIE, P., BRABANT, L., & VAN HOOREBEKE, L.
- Affiliations
- Ghent University (BE)
- Corresponding author
- PAUWELS, E.
- Venue
- Journal of Microscopy
- Publisher
- Wiley
- Published
- 2013-02-22
- Volume
- 250
- Issue
- 1
- Pages
- 21-31
- ISSN
- 0022-2720 (ISSN-L)
- 1365-2818
- OpenAlex
- W1833649979
- Cited by
- 212
- References
- 38
- Citations by year
- 2026: 14 · 2025: 12 · 2024: 15 · 2023: 13 · 2022: 17 · 2021: 14 · 2020: 23 · 2019: 28 · 2018: 24 · 2017: 20 · 2016: 10 · 2015: 8 · 2014: 9 · 2013: 4 · 2012: 1
- FWCI
- 10.23
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Microscopy (publishedVersion)
- PubMed (publishedVersion)
- Ghent University Academic Bibliography (Ghent University) (submittedVersion)
- Topics
- Physical Sciences › Physics and Astronomy › Radiation › Advanced X-ray Imaging Techniques
- Physical Sciences › Engineering › Biomedical Engineering › Advanced X-ray and CT Imaging
- Health Sciences › Medicine › Urology › Comparative Animal Anatomy Studies
- Keywords
- X-ray contrast agents, soft tissue visualization, micro-CT, contrast enhancement, tissue staining, mercuric chloride, phosphotungstic acid, phosphomolybdic acid, sodium tungstate, potassium iodide, penetration depth, tissue fixation
- MeSH terms
- Animals, Contrast Media, Image Processing, Computer-Assisted, Staining and Labeling, Swine, Lower Extremity, Mice, X-Ray Microtomography
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