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Abstract BackgroundGiven the need for descriptive and increasingly mechanistic morphological analyses, contrast‐enhanced microcomputed tomography (microCT) represents perhaps the best method for visualizing 3D biological soft tissues in situ. Although staining protocols using phosphotungstic acid (PTA) have been published with beautiful visualizations of soft tissue structures, these protocols are often aimed at highly specific research questions and are applicable to a limited set of model organisms, specimen ages, or tissue types. We provide detailed protocols for micro‐level visualization of soft tissue structures in mice at several embryonic and early postnatal ages using PTA‐enhanced microCT. ResultsOur protocols produce microCT scans that enable visualization and quantitative analyses of whole organisms, individual tissues, and organ systems while preserving 3D morphology and relationships with surrounding structures, with minimal soft tissue shrinkage. Of particular note, both internal and external features of the murine heart, lungs, and liver, as well as embryonic cartilage, are captured at high resolution. ConclusionThese protocols have broad applicability to mouse models for a variety of diseases and conditions. Minor experimentation in the staining duration can expand this protocol to additional age groups, permitting ontogenetic studies of internal organs and soft tissue structures within their 3D in situ position.
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Getting Nosy: Olfactory Rosette Morphology and Lamellar Microstructure of Two Chondrichthyan Species
Synopsis To smell, fish rely on passive water flow into their olfactory chambers and through their olfactory rosettes to detect chemical signals in their aquatic environment. The olfactory rosette is made up of secondarily folded tissues called olfactory lamellae. The olfactory morphology of cartilaginous fishes varies widely in both rosette gross morphology and lamellar microstructure. Previous research has shown differences in lamellar sensory morphology depending on the position along the rosette in hammerheads (family Sphyrnidae). Here, we investigate if this pattern continues in members of two other chondrichthyan families: Squalidae and Chimaeridae. Using contrast-enhanced microCT and scanning electron microscopy, we investigated patterns in lamellar morphology based on lamellar position along the olfactory rosette in Pacific spiny dogfish (Squalus suckleyi) and spotted ratfish (Hydrolagus colliei). We describe the gross olfactory rosette anatomy and lamellar microstructure of both species. We also put forth a new method, combining 3D morphological microCT data with 2D SEM microstructure data to better approximate lamellar sensory surface area. We found that in both species, lamellae in the center of the rosette were larger with more secondary folds. However, we found no significant differences in lamellar sensory surface area among lamellar positions. Previously, differences in lamellar sensory morphology have been tied to the internal fluid dynamics of the olfactory chamber. It is possible that the internal flow dynamics of these species are like other chondrichthyan models, where water flow patterns differ in the lateral vs the medial part of the organ, and the consistent distribution of sensory tissue does not correspond to this flow. Alternatively, the olfactory morphology of these species may result in uniform flow patterns throughout the olfactory chamber, correlating with the consistent distribution of sensory tissue throughout the organ. This study emphasizes that further investigations into chondrichthyan fluid dynamics is paramount to any future studies on the correlations between distribution of sensory tissues and water flow.
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- Award ID(s):
- 2109408
- PAR ID:
- 10542489
- Publisher / Repository:
- Oxford University Press
- Date Published:
- Journal Name:
- Integrative And Comparative Biology
- Volume:
- 64
- Issue:
- 2
- ISSN:
- 1540-7063
- Format(s):
- Medium: X Size: p. 441-458
- Size(s):
- p. 441-458
- Sponsoring Org:
- National Science Foundation
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