Three-dimensional observation of mouse tongue muscles using micro-computed tomography

Aoyagi, Hidekazu; Iwasaki, Shigeo; Nakamura, Kenzirou

doi:10.1007/s10266-013-0131-4

Cited by 12 publications

(18 citation statements)

References 35 publications

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“…So far no widespread application of µCT for the evaluation of soft tissues in a similar manner is routinely used due to the low X-ray attenuation of non-mineralized tissues. Commonly used contrast agents such as osmium or gold [5][6][7][8] bear the disadvantages of high cost or toxicity. To overcome this limitation, alternative staining methods including iodine-based dyes such as Lugol's solution were adapted for µCT use, as first described by Metscher [9,10].…”

Section: Introductionmentioning

confidence: 99%

Iodine-enhanced micro-CT imaging of soft tissue on the example of peripheral nerve regeneration

Heimel¹,

Swiadek

Slezak

et al. 2018

Preprint

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Microcomputed tomography (µCT) is widely used for the study of mineralized tissues but a similar use for soft tissues is hindered by their low X-ray attenuation. This limitation can be overcome by the recent development of different staining techniques. Staining with Lugol's solution, a mixture of one part iodine and two parts potassium iodide in water, stands out among these techniques for its low complexity and cost. Currently, Lugol staining is mostly used for anatomical examination of tissues. In the present study we seek to optimize the quality and reproducibility of the staining for ex vivo visualization of soft tissues in the context of a peripheral nerve regeneration model in the rat.We show that the staining result not only depends on the concentration of the staining solution, but also on the amount of stain in relation to the tissue volume and composition, necessitating careful adaptation of the staining protocol to the respective specimen tissue. This optimization can be simplified by a stepwise staining which we show to yield a similar result compared to staining in a single step. Lugol staining solution results in concentration dependent tissue shrinkage which can be minimized but not eliminated. We compared the shrinkage of tendon, nerve, skeletal muscle, heart, brain and kidney with six iterations of Lugol staining. 60 ml of 0.3% Lugol's solution per cm³ of tissue for 24h yielded good results on the example of a peripheral nerve regeneration model and we were able to show that the regenerating nerve inside a silk fibroin tube can be visualized in 3D using this staining technique. This information helps in deciding the region of interest for histological imaging and provides a 3D context to histological findings. Correlating both imaging modalities has the potential to improve the understanding of the regenerative process.

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Section: Introductionmentioning

confidence: 99%

Iodine-enhanced micro-CT imaging of soft tissue on the example of peripheral nerve regeneration

Heimel¹,

Swiadek

Slezak

et al. 2018

Preprint

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“…To be more specific, OsO 4 gives better resolution on lipids, proteins, and nucleic acids. Three-dimensional observation or murine tongue muscles using CT techniques was reported by Aoyagi, Iwasaki, and Nakamura (2015). Three-month-old mice head with tongues was fixed with 4% formalin for 1 week at room temperature and freezedried; then, the tongues were stained for 24 h in 1% OsO 4 at 4 C.…”

Section: Phosphomolybdic Acidmentioning

confidence: 99%

Evaluation of X‐ray tomography contrast agents: A review of production, protocols, and biological applications

Koç

Aslan

Kao

et al. 2019

Microscopy Res & Technique

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X‐ray computed tomography is a strong tool that finds many applications both in medical applications and in the investigation of biological and nonbiological samples. In the clinics, X‐ray tomography is widely used for diagnostic purposes whose three‐dimensional imaging in high resolution helps physicians to obtain detailed image of investigated regions. Researchers in biological sciences and engineering use X‐ray tomography because it is a nondestructive method to assess the structure of their samples. In both medical and biological applications, visualization of soft tissues and structures requires special treatment, in which special contrast agents are used. In this detailed report, molecule‐based and nanoparticle‐based contrast agents used in biological applications to enhance the image quality were compiled and reported. Special contrast agent applications and protocols to enhance the contrast for the biological applications and works to develop nanoparticle contrast agents to enhance the contrast for targeted drug delivery and general imaging applications were also assessed and listed.

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“…These metallic contrasting agents were developed for earlier imaging techniques, such as osmium tetroxide for electron transmission microscopy (Carson and Hladik, ) and Lugol's iodine (I 2 KI) for radiography and microdissection (Wallingford, ; Bock and Shear, ). However, osmium tetroxide is highly toxic, has a low penetration rate (Aoyagi et al, ), and is, therefore, used primarily for CT scanning of small objects, whereas Lugol's iodine is much less toxic, diffuses into tissues much more quickly than PMA or PTA, and has, therefore, been used preferentially for larger samples (Degenhardt et al, ; Pauwels et al, ; Descamps et al, ; Gignac et al, ). In general, higher concentrations of Lugol's iodine penetrate tissues more quickly (Gignac and Kley, ; Gignac et al, ), but they cause greater tissue shrinkage and distortion (Degenhardt et al, ; Pauwels et al, ; Vickerton et al, ; Wong et al, ; Buytaert et al, ; Descamps et al, ; Schmidbaur et al, ).…”

Section: Introductionmentioning

confidence: 99%

3DMRIModeling of Thin and Spatially Complex Soft Tissue Structures without Shrinkage: Lamprey Myosepta as an Example

Wood

Jia

Carmichael

et al. 2018

The Anatomical Record

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3D imaging techniques enable the nondestructive analysis and modeling of complex structures. Among these, MRI exhibits good soft tissue contrast, but is currently less commonly used for nonclinical research than X-ray CT, even though the latter requires contrast-staining that shrinks and distorts soft tissues. When the objective is the creation of a realistic and complete 3D model of soft tissue structures, MRI data are more demanding to acquire and visualize and require extensive post-processing because they comprise noncubic voxels with dimensions that represent a trade-off between tissue contrast and image resolution. Therefore, thin soft tissue structures with complex spatial configurations are not always visible in a single MRI dataset, so that standard segmentation techniques are not sufficient for their complete visualization. By using the example of the thin and spatially complex connective tissue myosepta in lampreys, we developed a workflow protocol for the selection of the appropriate parameters for the acquisition of MRI data and for the visualization and 3D modeling of soft tissue structures. This protocol includes a novel recursive segmentation technique for supplementing missing data in one dataset with data from another dataset to produce realistic and complete 3D models. Such 3D models are needed for the modeling of dynamic processes, such as the biomechanics of fish locomotion. However, our methodology is applicable to the visualization of any thin soft tissue structures with complex spatial configurations, such as fasciae, aponeuroses, and small blood vessels and nerves, for clinical research and the further exploration of tensegrity. Anat Rec, 301:1745-1763, 2018. © 2018 Wiley Periodicals, Inc.

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Three-dimensional observation of mouse tongue muscles using micro-computed tomography

Cited by 12 publications

References 35 publications

Iodine-enhanced micro-CT imaging of soft tissue on the example of peripheral nerve regeneration

Iodine-enhanced micro-CT imaging of soft tissue on the example of peripheral nerve regeneration

Evaluation of X‐ray tomography contrast agents: A review of production, protocols, and biological applications

3DMRIModeling of Thin and Spatially Complex Soft Tissue Structures without Shrinkage: Lamprey Myosepta as an Example

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