Diffusible iodine‐based contrast‐enhanced computed tomography (diceCT): an emerging tool for rapid, high‐resolution, 3‐D imaging of metazoan soft tissues
Open accessAbstract
Morphologists have historically had to rely on destructive procedures to visualize the three‐dimensional (3‐D) anatomy of animals. More recently, however, non‐destructive techniques have come to the forefront. These include X‐ray computed tomography ( CT ), which has been used most commonly to examine the mineralized, hard‐tissue anatomy of living and fossil metazoans. One relatively new and potentially transformative aspect of current CT ‐based research is the use of chemical agents to render visible, and differentiate between, soft‐tissue structures in X‐ray images. Specifically, iodine has emerged as one of the most widely used of these contrast agents among animal morphologists due to its ease of handling, cost effectiveness, and differential affinities for major types of soft tissues. The rapid adoption of iodine‐based contrast agents has resulted in a proliferation of distinct specimen preparations and scanning parameter choices, as well as an increasing variety of imaging hardware and software preferences. Here we provide a critical review of the recent contributions to iodine‐based, contrast‐enhanced CT research to enable researchers just beginning to employ contrast enhancement to make sense of this complex new landscape of methodologies. We provide a detailed summary of recent case studies, assess factors that govern success at each step of the specimen storage, preparation, and imaging processes, and make recommendations for standardizing both techniques and reporting practices. Finally, we discuss potential cutting‐edge applications of diffusible iodine‐based contrast‐enhanced computed tomography (dice CT ) and the issues that must still be overcome to facilitate the broader adoption of dice CT going forward.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
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 [18]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Humason, G. L. (1962) · W.H. Freeman eBooks · doi:10.5962/bhl.title.5890 · Cited by 3,657 · see citing works
Vogel's textbook of quantitative chemical analysis
Vogel, A. I., & Jeffery, G. H. (1989) · Medical Entomology and Zoology · Cited by 2,259 · see citing works
Ketcham, R. A., & Carlson, W. D. (2001) · Computers & Geosciences · doi:10.1016/s0098-3004(00)00116-3 · Cited by 1,537 · see citing works
The stabilization of proteins by sucrose.
Lee, J. C., & Timasheff, S. N. (1981) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)68947-7 · Cited by 1,112 · see citing works
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
Dunnill, M. S. (1973) · Journal of Clinical Pathology · doi:10.1136/jcp.26.4.316-b · Cited by 380 · see citing works
Deriving Hounsfield units using grey levels in cone beam computed tomography
Mah, P., Reeves, T., & McDavid, W. D. (2010) · Dentomaxillofacial Radiology · doi:10.1259/dmfr/19603304 · Cited by 372 · see citing works
MicroComputed Tomography: Methodology and Applications
Stock, S. R. (2008) · Thammasat University Digital Collections · Cited by 325 · 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
Gignac, P. M., & Kley, N. J. (2014) · Journal of Experimental Zoology Part B Molecular and Developmental Evolution · doi:10.1002/jez.b.22561 · Cited by 247 · see citing works
Self-citationFunctional Evolution of the Feeding System in Rodents
Cox, P. G., Rayfield, E. J., Fagan, M. J., et al. (2012) · PLoS ONE · doi:10.1371/journal.pone.0036299 · Cited by 243 · see citing works
Self-citationMicro-computed tomography: Introducing new dimensions to taxonomy
Faulwetter, S., Vasileiadou, K., Kouratoras, M., Dailianis, T., & Arvanitidis, C. (2013) · ZooKeys · doi:10.3897/zookeys.263.4261 · Cited by 228 · 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
Self-citationX-ray microtomography in biology
Mizutani, R., & Suzuki, Y. (2011) · Micron · doi:10.1016/j.micron.2011.10.002 · Cited by 218 · see citing works
Düring, D. N., Ziegler, A., Thompson, C. K., Ziegler, A., Faber, C., Müller, J., Scharff, C., & Elemans, C. P. H. (2013) · BMC Biology · doi:10.1186/1741-7007-11-1 · Cited by 213 · see citing works
Self-citationPAUWELS, E., VAN LOO, D., CORNILLIE, P., BRABANT, L., & VAN HOOREBEKE, L. (2013) · Journal of Microscopy · doi:10.1111/jmi.12013 · Cited by 212 · see citing works
Chemical demonstrations : a handbook for teachers of chemistry.
Shakhashiri, B. Z. (1983) · Medical Entomology and Zoology · Cited by 211 · see citing works
Concentration‐dependent specimen shrinkage in iodine‐enhanced micro CT
Vickerton, P., Jarvis, J. C., & Jeffery, N. S. (2013) · Journal of Anatomy · doi:10.1111/joa.12068 · Cited by 190 · see citing works
Buytaert, J., Goyens, J., Greef, D. D., Aerts, P., & Dirckx, J. (2014) · Microscopy and Microanalysis · doi:10.1017/s1431927614001329 · Cited by 174 · see citing works
Cox, P. G., & Jeffery, N. S. (2011) · The Anatomical Record · doi:10.1002/ar.21381 · Cited by 167 · see citing works
Self-citationThe morphology of the mouse masticatory musculature
Baverstock, H., Jeffery, N. S., & Cobb, S. N. (2013) · Journal of Anatomy · doi:10.1111/joa.12059 · Cited by 146 · see citing works
Sombke, A., Lipke, E., Michalik, P., Uhl, G., & Harzsch, S. (2015) · Journal of Comparative Neurology · doi:10.1002/cne.23741 · Cited by 144 · see citing works
Soft tissue discrimination with contrast agents using micro-CT scanning
Descamps, E., Sochacka, A., Kegel, B. D., et al. (2020) · Belgian journal of zoology · doi:10.26496/bjz.2014.63 · Cited by 144 · see citing works
A novel 3D mouse embryo atlas based on micro-CT
Wong, M. D., Dorr, A. E., Walls, J. R., Lerch, J. P., & Henkelman, R. M. (2012) · Development · doi:10.1242/dev.082016 · Cited by 141 · see citing works
Self-citationHautier, L., Lebrun, R., & Cox, P. G. (2012) · Journal of Morphology · doi:10.1002/jmor.20061 · Cited by 127 · see citing works
Self-citationStephenson, R. S., Boyett, M. R., Hart, G., Nikolaidou, T., Cai, X., Corno, A. F., Alphonso, N., Jeffery, N. S., & Jarvis, J. C. (2012) · PLoS ONE · doi:10.1371/journal.pone.0035299 · Cited by 124 · see citing works
Cooper, R. A. (2007) · International Wound Journal · doi:10.1111/j.1742-481x.2007.00314.x · Cited by 124 · see citing works
A 3D Interactive Model and Atlas of the Jaw Musculature of Alligator mississippiensis
Holliday, C. M., Tsai, H. P., Skiljan, R. J., et al. (2013) · PLoS ONE · doi:10.1371/journal.pone.0062806 · Cited by 122 · see citing works
Self-citationVogel's textbook of quantitative chemical analysis. 5th edn
Leonard, M. A. (1990) · Endeavour · doi:10.1016/0160-9327(90)90087-8 · Cited by 120 · see citing works
Kondo, H. G., Fukuda, H., Ono, H., et al. (2001) · Gastrointestinal Endoscopy · doi:10.1067/mge.2001.110730 · Cited by 118 · see citing works
Balanoff, A. M., Bever, G. S., Colbert, M. W., Clarke, J. A., Field, D. J., Gignac, P. M., Ksepka, D. T., Ridgely, R. C., Smith, N. A., Torres, C. R., et al. (2015) · Journal of Anatomy · doi:10.1111/joa.12378 · Cited by 118 · see citing works
Self-citationRelationship between Hounsfield unit in CT scan and gray scale in CBCT
Kamaruddin, N., Rajion, Z. A., Yusof, A., et al. (2016) · AIP conference proceedings · doi:10.1063/1.4968860 · Cited by 116 · see citing works
Linnaeus in the information age
Godfray, H. C. J. (2007) · Nature · doi:10.1038/446259a · Cited by 113 · see citing works
Relationship between Hounsfield Unit in CT Scan and Gray Scale in CBCT
Razi, T., Niknami, M., & Ghazani, F. A. (2014) · PubMed · doi:10.5681/joddd.2014.019 · Cited by 112 · see citing works
Aslanidi, O. V., Nikolaidou, T., Zhao, J., Smaill, B. H., Gilbert, S., Holden, A. V., Lowe, T., Withers, P. J., Stephenson, R. S., Jarvis, J. C., et al. (2012) · IEEE Transactions on Medical Imaging · doi:10.1109/tmi.2012.2209183 · Cited by 110 · see citing works
Three-dimensional non-destructive soft-tissue visualization with X-ray staining micro-tomography
Silva, J. M. D. S. E., Zanette, I., Noël, P. B., et al. (2015) · Scientific Reports · doi:10.1038/srep14088 · Cited by 105 · 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
Cox, P. G., Fagan, M. J., Rayfield, E. J., & Jeffery, N. S. (2011) · Journal of Anatomy · doi:10.1111/j.1469-7580.2011.01436.x · Cited by 101 · see citing works
Self-citationStructural Stabilization of Tissue for Embryo Phenotyping Using Micro-CT with Iodine Staining
Wong, M. D., Spring, S., & Henkelman, R. M. (2013) · PLoS ONE · doi:10.1371/journal.pone.0084321 · Cited by 94 · see citing works
Self-citationGeorge, I. D., & Holliday, C. M. (2013) · The Anatomical Record · doi:10.1002/ar.22666 · Cited by 93 · see citing works
Self-citationAkkari, N., Enghoff, H., & Metscher, B. (2015) · PLoS ONE · doi:10.1371/journal.pone.0135243 · Cited by 88 · see citing works
Artificial selection for high activity favors mighty mini-muscles in house mice
Houle-Leroy, P., Guderley, H., Swallow, J. G., et al. (2003) · American Journal of Physiology-Regulatory, Integrative and Comparative Physiology · doi:10.1152/ajpregu.00179.2002 · Cited by 88 · see citing works
Lautenschlager, S., Bright, J. A., & Rayfield, E. J. (2013) · Journal of Anatomy · doi:10.1111/joa.12153 · Cited by 83 · see citing works
Tsai, H. P., & Holliday, C. M. (2011) · PLoS ONE · doi:10.1371/journal.pone.0024935 · Cited by 81 · see citing works
Self-citationValiyaparambil, J. V., Yamany, I. A., Ortiz, D., et al. (2013) · PubMed · Cited by 81 · see citing works
Tsai, H. P., & Holliday, C. M. (2014) · Journal of Morphology · doi:10.1002/jmor.20360 · Cited by 75 · see citing works
Self-citationCox, P. G., & Faulkes, C. G. (2014) · PeerJ · doi:10.7717/peerj.448 · Cited by 75 · see citing works
Self-citationFernández, R., Kvist, S., Lenihan, J. W., et al. (2014) · PLoS ONE · doi:10.1371/journal.pone.0096617 · Cited by 73 · see citing works
Hutchinson, J. C., Arthurs, O. J., Ashworth, M. T., Ramsey, A. T., Mifsud, W., Lombardi, C. M., & Sebire, N. J. (2015) · Ultrasound in Obstetrics and Gynecology · doi:10.1002/uog.15764 · Cited by 69 · see citing works
Syme, D. A., Evashuk, K., Grintuch, B., et al. (2005) · Journal of Applied Physiology · doi:10.1152/japplphysiol.00369.2005 · Cited by 62 · see citing works
The Development of Organic Iodine Compounds as X-ray Contrast Media
Wallingford, V. H. (1953) · Journal of the American Pharmaceutical Association (Scientific ed ) · doi:10.1002/jps.3030421206 · Cited by 62 · see citing works
Automated pipeline for anatomical phenotyping of mouse embryos using micro-CT
Wong, M. D., Maezawa, Y., Lerch, J. P., & Henkelman, R. M. (2014) · Development · doi:10.1242/dev.107722 · Cited by 62 · see citing works
Self-citationDisplay and Visualization of Three-Dimensional Reconstructed Anatomic Morphology
Harris, L. D., Robb, R. A., Yuen, T. S., et al. (1979) · Journal of Computer Assisted Tomography · doi:10.1097/00004728-197908000-00002 · Cited by 61 · see citing works
Handbook of Imaging in Biological Mechanics
(2014) · CRC Press eBooks · doi:10.1201/b17566 · Cited by 52 · see citing works
Hopkins, T., Heilman, A. M., Liggett, J. A., et al. (2015) · Journal of Neuroscience Methods · doi:10.1016/j.jneumeth.2015.08.016 · Cited by 49 · see citing works
Testing hypotheses of bat baculum function with 3D models derived from microCT
Herdina, A. N., Kelly, D. A., Jahelková, H., et al. (2015) · Journal of Anatomy · doi:10.1111/joa.12274 · Cited by 47 · see citing works
Self-citationTahara, R., & Larsson, H. C. E. (2013) · Journal of Anatomy · doi:10.1111/joa.12081 · Cited by 46 · see citing works
Guderley, H., Houle-Leroy, P., Diffee, G. M., Camp, D. M., & Garland, T. (2006) · Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology · doi:10.1016/j.cbpb.2006.02.009 · Cited by 45 · see citing works
Experimental reduction of intromittent organ length reduces male reproductive success in a bug
Dougherty, L. R., Rahman, I. A., Burdfield‐Steel, E., et al. (2015) · Proceedings of the Royal Society B Biological Sciences · doi:10.1098/rspb.2015.0724 · Cited by 44 · see citing works
THE DETERMINATION OF IODINE IN BIOLOGICAL MATERIAL
Trevorrow, V. E., & Fashena, G. J. (1935) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)75149-1 · Cited by 41 · see citing works
Frog tongue acts as muscle-powered adhesive tape
Kleinteich, T., & Gorb, S. N. (2015) · Royal Society Open Science · doi:10.1098/rsos.150333 · Cited by 41 · see citing works
Phenotypic Effects of the "Mini-Muscle" Allele in a Large HR x C57BL/6J Mouse Backcross
Hannon, R. M., Kelly, S. A., Middleton, K. M., et al. (2008) · Journal of Heredity · doi:10.1093/jhered/esn011 · Cited by 38 · see citing works
Li, Z., & Clarke, J. A. (2015) · Evolutionary Biology · doi:10.1007/s11692-015-9345-4 · Cited by 36 · see citing works
Self-citationVickerton, P., Jarvis, J. C., Gallagher, J. A., et al. (2014) · Proceedings of the Royal Society B Biological Sciences · doi:10.1098/rspb.2014.0786 · Cited by 36 · see citing works
Metscher, B. (2013) · Cited by 34 · see citing works
A mouse informatics platform for phenotypic and translational discovery
Ring, N., Meehan, T. F., Blake, A., Brown, J. M., Chen, C.-K., Conte, N., Fenza, A. D., Fiegel, T., Horner, N., Jacobsen, J. O. B., et al. (2015) · Mammalian Genome · doi:10.1007/s00335-015-9599-2 · Cited by 32 · see citing works
Imaging modalities to assess structural birth defects in mutant mouse models
Tobita, K., Liu, X., & Lo, C. W. Y. (2010) · Birth Defects Research Part C Embryo Today Reviews · doi:10.1002/bdrc.20187 · Cited by 31 · see citing works
Herdina, A. N., Herzig‐Straschil, B., Hilgers, H., et al. (2010) · The Anatomical Record · doi:10.1002/ar.21148 · Cited by 27 · see citing works
Self-citationHerdina, A. N., Plenk, H., Jr., Benda, P., et al. (2015) · Journal of Morphology · doi:10.1002/jmor.20372 · Cited by 24 · see citing works
Self-citationJaspers, C., & Carstensen, J. (2009) · Limnology and Oceanography Methods · doi:10.4319/lom.2009.7.430 · Cited by 22 · see citing works
Lenihan, J. W., Kvist, S., Fernández, R., et al. (2014) · GigaScience · doi:10.1186/2047-217x-3-6 · Cited by 18 · see citing works
Fundamentals of Body CT, 3rd ed
Baumgarten, D. A. (2006) · American Journal of Roentgenology · doi:10.2214/ajr.06.5074 · Cited by 4 · see citing works
Spaw, A., & Witmer, L. M. (2014) · The FASEB Journal · doi:10.1096/fasebj.28.1_supplement.923.6 · Cited by 2 · see citing works
Self-citationSpaw, A., & Witmer, L. M. (2013) · The FASEB Journal · doi:10.1096/fasebj.27.1_supplement.531.4 · Cited by 2 · see citing works
Self-citation
Flagged first, then by citations
Research works printed but not linked: 1
- Vickerton P, Jarvis J, Jeffery NS (2013) Concentration‐dependent specimen shrinkage in iodine‐enhanced microCT. J Anat 223, 185– 193. 23721431 PMC3724211
Cited by
Sorted by citations
Open data and digital morphology
Davies, T. G., Rahman, I. A., Lautenschlager, S., Cunningham, J. A., Asher, R. J., Barrett, P. M., Bates, K. T., Bengtson, S., Benson, R. B. J., Boyer, D. M., et al. (2017) · Proceedings of the Royal Society B: Biological Sciences · doi:10.1098/rspb.2017.0194 · Cited by 181 · see citing works
Self-citationHANDSCHUH, S., BEISSER, C. J., RUTHENSTEINER, B., & METSCHER, B. D. (2017) · Journal of Microscopy · doi:10.1111/jmi.12543 · Cited by 48 · see citing works
High-throughput phenomics of global ant biodiversity
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
- Authors
- Gignac, P. M., Kley, N. J., Clarke, J. A., Colbert, M. W., Morhardt, A. C., Cerio, D., Cost, I. N., Cox, P. G., Daza, J. D., Early, C. M., Echols, M. S., Henkelman, R. M., Herdina, A. N., Holliday, C. M., Li, Z., Mahlow, K., Merchant, S., Müller, J., Orsbon, C. P., Paluh, D. J., Thies, M. L., Tsai, H. P., & Witmer, L. M.
- Affiliations
- Oklahoma State University Center for Health Sciences (US)
- Stony Brook University (US)
- The University of Texas at Austin (US)
- Ohio University (US)
- University of Missouri (US)
- University of York (GB)
- Sam Houston State University (US)
- OAK Orthopedics (US)
- University of Toronto (CA)
- University of Vienna (AT)
- Museum für Naturkunde (DE)
- University of Utah (US)
- University of Chicago (US)
- Villanova University (US)
- Brown University (US)
- Corresponding author
- Gignac, Paul M.
- ORCID
- Gignac, Paul M.
- Clarke, Julia A.
- Colbert, Matthew W.
- Morhardt, Ashley C.
- Cerio, Donald
- Cost, Ian N.
- Cox, Philip G.
- Daza, Juan D.
- Early, Catherine M.
- Henkelman, R. Mark
- Herdina, A. Nele
- Holliday, Casey M.
- Li, Zhiheng
- Mahlow, Kristin
- Merchant, Samer
- Müller, Johannes
- Orsbon, Courtney P.
- Paluh, Daniel J.
- Tsai, Henry P.
- Witmer, Lawrence M.
- Venue
- Journal of Anatomy
- Publisher
- Wiley
- Published
- 2016-03-11
- Volume
- 228
- Issue
- 6
- Pages
- 889-909
- ISSN
- 0021-8782 (ISSN-L)
- 1469-7580
- PMC
- PMC5341577
- OpenAlex
- W2296094736
- Cited by
- 541
- References
- 75
- Citations by year
- 2026: 36 · 2025: 49 · 2024: 63 · 2023: 60 · 2022: 48 · 2021: 65 · 2020: 62 · 2019: 59 · 2018: 46 · 2017: 37 · 2016: 11 · 2013: 1 · 2012: 1
- FWCI
- 11.26
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Anatomy (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- White Rose Research Online (University of Leeds, The University of Sheffield, University of York) (acceptedVersion)
- BearWorks (Missouri State University) (submittedVersion)
- Funders
- National Science Foundation (NSF IOS‐0749750, NSF EAGER 1450842, NSF DEB‐1457180, NSF EAGER 1450850)
- Topics
- Health Sciences › Medicine › Urology › Comparative Animal Anatomy Studies
- Physical Sciences › Physics and Astronomy › Radiation › Advanced X-ray Imaging Techniques
- Physical Sciences › Engineering › Biomedical Engineering › Anatomy and Medical Technology
- Keywords
- diceCT, iodinated contrast media, contrast-enhanced CT, soft tissue imaging, specimen preparation, specimen storage, 3D imaging, non-destructive imaging
- MeSH terms
- Anatomy, Comparative, Animals, Contrast Media, Iodides, Tomography, X-Ray Computed, Imaging, Three-Dimensional
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
