Abstract
The mammalian cochlea is a complex macroscopic structure due to its helical shape and the microscopic arrangements of the individual layers of cells. To improve the outcomes of hearing restoration in deaf patients, it is important to understand the anatomic structure and composition of the cochlea ex vivo. Hitherto, only one histological technique based on confocal laser scanning microscopy and optical clearing has been developed for in toto optical imaging of the murine cochlea. However, with a growing size of the specimen, e.g., human cochlea, this technique reaches its limitations. Here, we demonstrate scanning laser optical tomography (SLOT) as a valuable imaging technique to visualize the murine cochlea in toto without any physical slicing. This technique can also be applied in larger specimens up to cm3 such as the human cochlea. Furthermore, immunolabeling allows visualization of inner hair cells (otoferlin) or spiral ganglion cells (neurofilament) within the whole cochlea. After image reconstruction, the 3D dataset was used for digital segmentation of the labeled region. As a result, quantitative analysis of position, length and curvature of the labeled region was possible. This is of high interest in order to understand the interaction of cochlear implants (CI) and cells in more detail.
Highlights
The anatomic structure of the cochlea is of high interest for efforts to improve the outcomes of hearing restoration
We show the capability of scanning laser optical tomography (SLOT) to image labeled regions inside the murine cochlea in toto, without the need for physical slicing of the sample
In the labeled sample 1, the spiral structure is visible in the maximum intensity projection (MIP)
Summary
The anatomic structure of the cochlea is of high interest for efforts to improve the outcomes of hearing restoration. The ex vivo visualization of the mammalian cochlea can be achieved by several imaging techniques. A widespread technique for ex vivo imaging is serial histological sectioning. The sample is immobilized by embedding in paraffin or by freezing for cryosections and cut in sections with a constant thickness [1]. These sections are mounted on microscope slides and can be visualized using 2D imaging. 3-dimensional registration of these images can be performed to generate a volumetric dataset of the cochlea Additional stains are applied to the sections, such as hematoxylin and eosin [2]. 3-dimensional registration of these images can be performed to generate a volumetric dataset of the cochlea
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