Iodine-enhanced micro-CT imaging: Methodological refinements for the study of the soft-tissue anatomy of post-embryonic vertebrates

Paul Gignac, Nathan J. Kley

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

The now widespread use of non-destructive X-ray computed tomography (CT) and micro-CT (μCT) has greatly augmented our ability to comprehensively detail and quantify the internal hard-tissue anatomy of vertebrates. However, the utility of X-ray imaging for gaining similar insights into vertebrate soft-tissue anatomy has yet to be fully realized due to the naturally low X-ray absorption of non-mineralized tissues. In this study, we show how a wide diversity of soft-tissue structures within the vertebrate head-including muscles, glands, fat deposits, perichondria, dural venous sinuses, white and gray matter of the brain, as well as cranial nerves and associated ganglia-can be rapidly visualized in their natural relationships with extraordinary levels of detail using iodine-enhanced (i-e) μCT imaging. To date, Lugol's iodine solution (I2KI) has been used as a contrast agent for μCT imaging of small invertebrates, vertebrate embryos, and certain isolated parts of larger, post-embryonic vertebrates. These previous studies have all yielded promising results, but visualization of soft tissues in smaller invertebrate and embryonic vertebrate specimens has generally been more complete than that for larger, post-embryonic vertebrates. Our research builds on these previous studies by using high-energy μCT together with more highly concentrated I2KI solutions and longer staining times to optimize the imaging and differentiation of soft tissues within the heads of post-embryonic archosaurs (Alligator mississippiensis and Dromaius novaehollandiae). We systematically quantify the intensities of tissue staining, demonstrate the range of anatomical structures that can be visualized, and generate a partial three-dimensional reconstruction of alligator cephalic soft-tissue anatomy.

Original languageEnglish
Pages (from-to)166-176
Number of pages11
JournalJournal of Experimental Zoology Part B: Molecular and Developmental Evolution
Volume322
Issue number3
DOIs
StatePublished - 1 Jan 2014

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micro-computed tomography
iodine
Iodine
anatomy
tomography
Vertebrates
Anatomy
vertebrate
Tomography
vertebrates
image analysis
computed tomography
Alligators and Crocodiles
X-radiation
Head
Invertebrates
Dromaiidae
invertebrate
invertebrates
X-Rays

Cite this

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abstract = "The now widespread use of non-destructive X-ray computed tomography (CT) and micro-CT (μCT) has greatly augmented our ability to comprehensively detail and quantify the internal hard-tissue anatomy of vertebrates. However, the utility of X-ray imaging for gaining similar insights into vertebrate soft-tissue anatomy has yet to be fully realized due to the naturally low X-ray absorption of non-mineralized tissues. In this study, we show how a wide diversity of soft-tissue structures within the vertebrate head-including muscles, glands, fat deposits, perichondria, dural venous sinuses, white and gray matter of the brain, as well as cranial nerves and associated ganglia-can be rapidly visualized in their natural relationships with extraordinary levels of detail using iodine-enhanced (i-e) μCT imaging. To date, Lugol's iodine solution (I2KI) has been used as a contrast agent for μCT imaging of small invertebrates, vertebrate embryos, and certain isolated parts of larger, post-embryonic vertebrates. These previous studies have all yielded promising results, but visualization of soft tissues in smaller invertebrate and embryonic vertebrate specimens has generally been more complete than that for larger, post-embryonic vertebrates. Our research builds on these previous studies by using high-energy μCT together with more highly concentrated I2KI solutions and longer staining times to optimize the imaging and differentiation of soft tissues within the heads of post-embryonic archosaurs (Alligator mississippiensis and Dromaius novaehollandiae). We systematically quantify the intensities of tissue staining, demonstrate the range of anatomical structures that can be visualized, and generate a partial three-dimensional reconstruction of alligator cephalic soft-tissue anatomy.",
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