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Assessing Soft-Tissue Shrinkage Estimates in Museum Specimens Imaged With Diffusible Iodine-Based Contrast-Enhanced Computed Tomography (diceCT)

Published online by Cambridge University Press:  19 June 2018

Brandon P. Hedrick*
Affiliation:
Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, MA 02138, USA
Laurel Yohe
Affiliation:
Department of Ecology and Evolution, Stony Brook University, 650 Life Sciences Building, Stony Brook, NY 11794, USA
Abby Vander Linden
Affiliation:
Graduate Program in Organismic and Evolutionary Biology, University of Massachusetts Amherst, Amherst, MA 01003, USA
Liliana M. Dávalos
Affiliation:
Department of Ecology and Evolution, Stony Brook University, 650 Life Sciences Building, Stony Brook, NY 11794, USA
Karen Sears
Affiliation:
Department of Animal Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
Alexa Sadier
Affiliation:
Department of Animal Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
Stephen J. Rossiter
Affiliation:
School of Biological and Chemical Sciences, Queen Mary University of London, London E1 4NS, UK
Kalina T. J. Davies
Affiliation:
School of Biological and Chemical Sciences, Queen Mary University of London, London E1 4NS, UK
Elizabeth Dumont
Affiliation:
School of Natural Sciences, University of California–Merced, Merced, CA 95343, USA
*
*Author for correspondence: Brandon P. Hedrick, E-mail: bphedrick1@gmail.com
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Abstract

The increased accessibility of soft-tissue data through diffusible iodine-based contrast-enhanced computed tomography (diceCT) enables comparative biologists to increase the taxonomic breadth of their studies with museum specimens. However, it is still unclear how soft-tissue measurements from preserved specimens reflect values from freshly collected specimens and whether diceCT preparation may affect these measurements. Here, we document and evaluate the accuracy of diceCT in museum specimens based on the soft-tissue reconstructions of brains and eyes of five bats. Based on proxies, both brains and eyes were roughly 60% of the estimated original sizes when first imaged. However, these structures did not further shrink significantly over a 4-week staining interval, and 1 week in 2.5% iodine-based solution yielded sufficient contrast for differentiating among soft-tissues. Compared to six “fresh” bat specimens imaged shortly after field collection (not fixed in ethanol), the museum specimens had significantly lower relative volumes of the eyes and brains. Variation in field preparation techniques and conditions, and long-term storage in ethanol may be the primary causes of shrinkage in museum specimens rather than diceCT staining methodology. Identifying reliable tissue-specific correction factors to adjust for the shrinkage now documented in museum specimens requires future work with larger samples.

Type
Biological Science Applications
Copyright
© Microscopy Society of America 2018 

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Footnotes

Cite this article: Hedrick BP, Yohe L, Vander Linden A, Dávalos LM, Sears K, Sadier A, Rossiter SJ, Davies KTJ, Dumont E (2018) Assessing Soft-Tissue Shrinkage Estimates in Museum Specimens Imaged With Diffusible Iodine-Based Contrast-Enhanced Computed Tomography (diceCT). Microsc Microanal24(3): 284–291. doi: 10.1017/S1431927618000399

References

Alvarez, J, Willig, MR, Jones, JK Webster, WD (1991) Glossophaga soricina. Mamm Species 379, 17.Google Scholar
Aslanidi, OV, Colman, MA, Zhao, J, Smaill, BH, Gilbert, SH, Hancox, JC, Boyett, MR Zhang, H (2012) Arrhythmogenic substrate for atrial fibrillation: Insights from an integrative computation model of pulmonary veins. 34th Ann Int Conf IEEE EMBS 28, 203206.Google Scholar
Aslanidi, OV, Nikolaidou, T, Zhao, J, Smaill, BH, Gilbert, SH, Holden, AV, Lowe, T, Withers, PJ, Stephenson, RS, Jarvis, JC, Hancox, JC, Boyett, MR Zhang, H (2013) Application of micro-computed tomography with iodine staining to cardiac imaging, segmentation, and computational model development. IEEE Trans Med Imaging 32, 817.CrossRefGoogle ScholarPubMed
Baverstock, H, Jeffery, NS Cobb, SN (2013) The morphology of the mouse masticatory musculature. J Anat 223, 4660.Google Scholar
Buytaert, J, Goyens, J, de Greef, D, Aerts, P Dirckx, J (2014) Volume shrinkage of bone, brain and muscle tissue in sample preparation for micro-CT and light sheet fluorescence microscopy (LSFM). Microsc Microanal 20, 12081217.Google Scholar
Chandler, N, Aslanidi, O, Buckley, D, Inada, S, Birchall, S, Atkinson, A, Kirk, D, Monfredi, O, Molenaar, P, Anderson, R, Shama, V, Sigg, D, Zhang, H, Boyett, M Dobrzynski, H (2011) Computer three-dimensional anatomical reconstruction of the human sinus node and a novel paranodal area. Anat Rec 294, 970979.Google Scholar
Cloutier, D Thomas, DW (1992) Carollia perspicillata . Mamm Species 417, 19.Google Scholar
Cox, PG Faulkes, CG (2014) Digital dissection of the masticatory muscles of the naked mole-rat, Heterocephalus glaber (Mammalia, Rodentia). Peer J 2, e448.Google Scholar
Cox, PG Jeffery, NS (2011) Reviewing the morphology of the jaw-closing musculature in squirrels, rats, and guinea pigs with contrast-enhanced microCT. Anat Rec 294, 915928.CrossRefGoogle ScholarPubMed
Da Cunha Tavares, V Mancina, CA (2008) Phyllops falcatus (Chiroptera: Phyllostomidae). Mamm Species 811, 17.Google Scholar
Dam, AN (1979) Shrinkage of the brain during histological procedures with fixation in formaldehyde solutions of different concentrations. J Hirnforsch 20, 115119.Google Scholar
Degenhardt, K, Wright, AC, Horng, D, Padmanabhan, A Epstein, JA (2010) Rapid 3D phenotyping of cardiovascular development in mouse embryos by microCT with iodine staining. Circ Cardiovasc Imaging 3, 314322.Google Scholar
Descamps, E, Sochacka, A, de Kegel, B, Van Loo, D, Van Hoorebeke, L Adriaens, D (2014) Soft-tissue discrimination with contrast agents using micro-CT scanning. Belg J Zool 144, 2040.Google Scholar
Düring, DN, Ziegler, A, Thompson, CK, Ziegler, A, Faber, C, Müller, J, Scharff, C Elemans, CPH (2013) The songbird syrinx morphome: A three-dimensional, high-resolution, interactive morphological map of the zebra finch vocal organ. BMC Biol 11, 127.Google Scholar
Ericsson, JLE Biberfeld, P (1967) Studies on aldehyde fixation. Lab Invest 17, 281.Google Scholar
Fox, CH, Johnson, FB, Whiting, J Roller, PP (1985) Formaldehyde fixation. J Histochem Cytochem 33, 845853.Google Scholar
George, ID Holliday, CM (2013) Trigeminal nerve morphology in Alligator mississippiensis and its significance for crocodyliform facial sensation and evolution. Anat Rec 296, 670680.CrossRefGoogle ScholarPubMed
Gignac, PM Kley, NJ (2014) Iodine-enhanced microCT imaging: Methodological refinements for the study of the soft-tissue anatomy of post-embryonic vertebrates. J Exp Zool 322B, 166176.CrossRefGoogle Scholar
Gignac, PM, Kley, NJ, Clarke, JA, Colbert, MW, Morhardt, AC, Cerio, D, Cost, IN, Cox, PG, Daza, JD, Early, CM, Echols, S, Henkelman, RM, Herdina, AN, Holliday, CM, Li, Z, Mahlow, K, Merchant, S, Müller, J, Orsbon, CP, Paluh, DJ, Thies, ML, Tsai, HP Witmer, LM (2016) Diffusible iodine-based contrast-enhanced computed tomography (diceCT): An emerging tool for rapid, high-resolution, 3-D imaging of metazoan soft tissues. J Anat 228, 889909.Google Scholar
Girard, R, Zeineddine, HA, Orsbon, C, Tan, H, Moore, T, Hobson, N, Shenkar, R, Lightle, R, Shi, C, Fam, MD, Cao, Y, Shen, L, Neander, AI, Rorrer, A, Gallione, C, Tang, AT, Kahn, ML, Marchuk, DA, Luo, Z Awad, IA (2016) Micro-computed tomography in murine models of cerebral cavernous malformations as a paradigm for brain disease. J Neurosci Methods 271, 1424.Google Scholar
Gold, MEL, Schulz, D, Budassi, M, Gignac, PM, Vaska, P Norell, MA (2016) Flying starlings, PET, and the evolution of volant dinosaurs. Curr Biol 26, R265R267.Google Scholar
Greenhall, AM, Joermann, G Schmidt, U (1983) Desmodus rotundus . Mamm Species 202, 16.Google Scholar
Hautier, L, Lebrun, R Cox, PG (2012) Patterns of covariation in the masticatory apparatus of hystricognathous rodents: Implications for evolution and diversification. J Morphol 273, 13191337.Google Scholar
Herdina, AN, Herzig-Straschil, B, Hilgers, H, Metscher, BD Plenk, H (2010) Histomorphology of the penis bone (baculum) in the Gray Long-Eared Bat Plecotus austriacus (Chiroptera, Vespertilionidae). Anat Rec 293, 12481258.Google Scholar
Herdina, AN, Kelly, DA, Jahelková, H, Lina, PHC, Horáček, I Metscher, BD (2015a) Testing hypotheses of bat baculum function with 3D models derived from microCT. J Anat 3, 229235.CrossRefGoogle Scholar
Herdina, AN, Plenk, H, Benda, P, Lina, PHC, Herzig-Straschil, B, Hilgers, H Metscher, BD (2015b) Correlative 3D-imaging of Pipistrellus penis micromorphology: Validating quantitative microCT images with undecalcified serial ground section histomorphology. J Morphol 276, 695706.Google Scholar
Hughes, DF, Walker, EM, Gignac, PM, Martinez, A, Negishi, K, Lieb, CS, Greenbaum, E Khan, AM (2016) Rescuing perishable neuroanatomical information from a threatened biodiversity hotspot: Remote field methods for brain tissue preservation validated by cytoarchitectonic analysis, immunohistochemistry, and x-ray microcomputed tomography. PLoS ONE 11(5), e0155824.Google Scholar
Jeffery, NS, Stephenson, RS, Gallagher, JA, Jarvis, JC Cox, PG (2011) Micro-computed tomography with iodine staining resolves the arrangement of muscle fibres. J Biomech 44, 189192.Google Scholar
Jennings, JB, Best, TL, Rainey, JC Burnett, SE (2000) Molossus pretiosus . Mamm Species 635, 13.Google Scholar
Li, Z, Ketchum, RA, Yan, F, Maisano, JA Clarke, JA (2016) Comparison and evaluation of the effectiveness of two approaches of diffusible iodine-based contrast enhanced computed tomography (diceCT) for avian cephalic material. J Exp Zool 326B, 352362.Google Scholar
Metscher, BD (2009 a) MicroCT for comparative morphology: Simple staining methods allow high-contrast 3D imaging of diverse non-mineralized animal tissues. BMC Physiol 9, 114.Google Scholar
Metscher, BD (2009 b) MicroCT for developmental biology: A versatile tool for high-contrast 3D imaging at histological resolutions. Dev Dyn 238, 632640.Google Scholar
Ortega, J Alarcón-D, I (2008) Anoura geoffroyi (Chiroptera: Phyllostomidae). Mamm Species 818, 17.Google Scholar
Ortega, J Castro-Arellano, I (2001) Artibeus jamaicensis . Mamm Species 662, 19.Google Scholar
Pauwels, E, Van Loo, D, Cornillie, P, Brabant, L Van Hoorebeke, L (2013) An exploratory study of contrast agents for soft tissue visualization by means of high resolution X-ray computed tomography imaging. J Microsc 250, 2131.Google Scholar
R Core Team (2016) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Available at https://www.R-project.org/.Google Scholar
Rehorek, SJ, Smith, TD Bhatnagar, KP (2010) The orbitofacial glands of bats: An investigation of the potential correlation of gland structure with social organization. Anat Rec 293, 14331448.Google Scholar
Rinehart, JB Kunz, TH (2006) Rhinophylla pumilio . Mamm Species 791, 15.Google Scholar
Ruff, S Wilson, D (1999) The Smithsonian Book of North American Mammals. Washington, DC: Smithsonian Institution Press. Pp. 1–750.Google Scholar
Schmidt, E, Parsons, T, Jamniczky, H, Gitelman, J, Trpkov, C, Boughner, JC, Logan, CC, Sensen, CW Hallgrímsson, B (2010) Micro-computed tomography-based phenotypic approaches in embryology: Procedural artifacts on assessments of embryonic craniofacial growth and development. BMC Dev Biol 10, 18.Google Scholar
Shenkar, R, Venkatasubramanian, PN, Wyrwicz, AM, Zhao, J, Shi, C, Akers, A, Marchuk, DA Awad, IA (2008) Advanced magnetic resonance imaging of cerebral cavernous malformations: II: Imaging of lesions in murine models. Neurosurgery 63, 790798.Google Scholar
Stephenson, RS, Boyett, MR, Hart, G, Nikolaidou, T, Cai, X, Corno, AF, Alphonso, N, Jeffery, NS Jarvis, JC (2012) Contrast enhanced micro-computed tomography resolves the 3-dimensional morphology of the cardiac conduction system in mammalian hearts. PLoS ONE 7(4), e35299.Google Scholar
Stockwell, E (2001) Morphology and flight manoeuvrability in new world leaf-nosed bats (Chiroptera: Phyllostomidae). J Zool 254, 505514.Google Scholar
Sturgess, JA Nicola, SJ (1975) Preparation of fish for identification and preservation as museum species. The Resources Agency of California, Department of Fish and Game, Inland Fisheries Informational Leaflet 29, Sacramento.Google Scholar
Tahara, R Larsson, HCE (2013) Quantitative analysis of microscopic x-ray computed tomography imaging: Japanese quail embryonic soft tissues with iodine staining. J Anat 223, 297310.Google Scholar
Tessler, M, Barrio, A, Borda, E, Rood-Goldman, R, Hill, M Siddall, ME (2016) Description of a soft-bodied invertebrate with microcomputed tomography and revision of the genus Chtonobdella (Hirudinea: Haemadipsidae). Zool Scr 45, 552565.Google Scholar
Tsai, HP Holliday, CM (2011) Ontogeny of the alligator cartilago transiliens and its significance for sauropsid jaw muscle evolution. PLoS ONE 6(9), e24935.Google Scholar
Vervust, B, Van Dongen, S Van Damme, R (2009) The effect of preservation on lizard morphometrics – An experimental study. Amphib Reptil 30, 321329.Google Scholar
Vickerton, P, Jarvis, JC Jeffery, NS (2013) Concentration-dependent specimen shrinkage in iodine-enhanced microCT. J Anat 223, 185193.Google Scholar
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