Abstract
Objectives/Hypothesis: In animal studies of vocal fold scarring and treatment, imaging-based evaluation is most often conducted by tissue slicing and histological staining. Given variation in anatomy, injury type, severity, and sacrifice timepoints, planar histological sections provide limited spatiotemporal details of tissue repair. Three-dimensional (3D) virtual histology may provide additional contextual spatial information, enhancing objective interpretation. The study's aim was to evaluate the suitability of magnetic resonance imaging (MRI), microscale computed tomography (CT), and nonlinear laser-scanning microscopy (NM) as virtual histology approaches for rabbit studies of vocal fold scarring. Methods: A unilateral injury was created using microcup forceps in the left vocal fold of three New Zealand White rabbits. Animals were sacrificed at 3, 10, and 39 days postinjury. ex vivo imaging of excised larynges was performed with MRI, CT, and NM modalities. Results: The MRI modality allowed visualization of injury location and morphological internal features with 100-μm spatial resolution. The CT modality provided a view of the injury defect surface with 12-μm spatial resolution. The NM modality with optical clearing resolved second-harmonic generation signal of collagen fibers and two-photon autofluorescence in vocal fold lamina propria, muscle, and surrounding cartilage structures at submicrometer spatial scales. Conclusions: Features of vocal fold injury and wound healing were observed with MRI, CT, and NM. The MRI and CT modalities provided contextual spatial information and dissection guidance, whereas NM resolved extracellular matrix structure. The results serve as a proof of concept to motivate incorporation of 3D virtual histology techniques in future vocal fold injury animal studies. Level of Evidence: NA Laryngoscope, 131:1578–1587, 2021.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1578-1587 |
| Number of pages | 10 |
| Journal | Laryngoscope |
| Volume | 131 |
| Issue number | 7 |
| DOIs | |
| State | Published - Jul 2021 |
| Externally published | Yes |
Bibliographical note
Funding Information:The authors acknowledge Melina Jaramillo Garcia for operation of the FVMPE-RS microscope at the Douglas Research Centre's Molecular and Cellular Microscopy Platform, an Olympus Discovery Center; Stephen Nuara for animal handling and operation of the Vimago platform, and Karen Hope Stone for animal study assistance and animal handling, at the Comparative Medicine and Animal Resources Center; Robert Gagnon at the Center for the Physics of Materials for aluminum sputter coating of glass slides; the McGill Physics Department MakerSpace for 3D printer access; and David Rudko for editing of the MRI methodology and discussion sections.
Funding Information:
This work was supported by the National Institute for Deafness and Other Communication Disorders of the National Institutes of Health under award number R01DC005788 and Natural Sciences and Engineering Research Council under award number RGPIN‐2018‐03843.
Publisher Copyright:
© 2020 American Laryngological, Rhinological and Otological Society Inc, "The Triological Society" and American Laryngological Association (ALA)
Keywords
- computed tomography
- magnetic resonance imaging
- multimodal imaging
- nonlinear microscopy
- Vocal fold scarring
PubMed: MeSH publication types
- Journal Article
- Research Support, N.I.H., Extramural
- Research Support, Non-U.S. Gov't