Use of micro computed-tomography and 3D printing for reverse engineering of mouse embryo nasal capsule

Abstract

Imaging of increasingly complex cartilage in vertebrate embryos is one of the key tasks of developmental biology. This is especially important to study shape-organizing processes during initial skeletal formation and growth. Advanced imaging techniques that are reflecting biological needs give a powerful impulse to push the boundaries of biological visualization. Recently, techniques for contrasting tissues and organs have improved considerably, extending traditional 2D imaging approaches to 3D. X-ray micro computed tomography (µCT), which allows 3D imaging of biological objects including their internal structures with a resolution in the micrometer range, in combination with contrasting techniques seems to be the most suitable approach for non-destructive imaging of embryonic developing cartilage. Despite there are many software-based ways for visualization of 3D data sets, having a real solid model of the studied object might give novel opportunities to fully understand the shape-organizing processes in the developing body. In this feasibility study we demonstrated the full procedure of creating a real 3D object of mouse embryo nasal capsule, i.e. the staining, the µCT scanning combined by the advanced data processing and the 3D printing. K : Computerized Tomography (CT) and Computed Radiography (CR); Image reconstruction in medical imaging; Multi-modality systems 1Corresponding author. Content from this work may be used under the terms of the Creative Commons Attribution 3.0 License.

Phosphotungstic acid‐enhanced microCT: Optimized protocols for embryonic and early postnatal mice

“…Analysis of the early development of the chondrocranium and its association with the forming skull has been hampered by the difficulty in visualizing embryonic cartilage in 3D. The recent Kaucka and Tesařová protocols demonstrated that PTA staining could successfully be used to visualize and analyze the chondrocranium, both in its entirety and with specific focus on the nasal capsule, in embryonic mice . Our expansion of the Kaucka and Tesařová protocols allows the segmentation of bone from the same PTA‐enhanced microCT scan, allowing the creation of high‐resolution 3D models of the forming dermatocranium and chondrocranium (Figure ).…”

“…Furthermore, the majority of published protocols provide only minimal details, requiring extensive experimentation to reach an optimized protocol, particularly if using a different model organism, specimen age, or tissue type. One research group has published detailed PTA protocols for use in embryonic mice from E12.5 to 18.5 . The Kaucka and Tesařová protocols added several significant steps to Metscher's original PTA staining method: a slow graded ethanol and methanol dehydration series, different suggested staining times for several ages of embryonic mice, and a final graded methanol rehydration series.…”

“…The slow dehydration and rehydration steps were added to help minimize soft tissue shrinkage, addressing many of the concerns with iodine staining. With these modifications, the Kaucka and Tesařová protocols provided excellent contrast enhancement, particularly for embryonic murine cranial soft tissues and cartilage . However, these protocols vary between publications, making it difficult to elect an optimum method.…”

“…However, these protocols vary between publications, making it difficult to elect an optimum method. For example, E15.5 embryos were either stained for 6 days with 0.7% PTA solution or 3 weeks with 1.0% to 1.5% PTA solution . These differences may reflect differences in the specific research goals for the stained specimens.…”

Three-dimensional imaging of internal tooth structures: Applications in dental education

“…As a remedy, 3D virtual models can now also be made accessible to dental students as custom-made and scaled physical objects through additive manufacturing (AM), thus providing students with a genuinely 3D interpretive experience [106,107]. AM is a process by which different types of material can be added layer-by-layer to produce a 3D object [108] and is increasingly being used in the life sciences [109][110][111].…”