A new approach to visualizing the membranous structures of the inner ear – high resolution X-ray micro-tomography

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Abstract
Through the application of high resolution X-ray micro-tomography and a method of contrast enhancement based on en bloc staining in osmium tetroxide (OsO4), we report an approach that facilitates accurate three-dimensional (3D) reconstruction to reveal the fine structure of the inner ear. To overcome the problems of artefacts, including tissue distortion and loss of 3D context that are inherent in existing methods that rely on manual dissection and/or histological sectioning. A staining protocol was developed that involved the en bloc application of the OsO4 solution (2% w/v) for an extended period of time. The samples were then scanned using an X-ray micro-tomography platform and subsequent 3D visualizations were constructed. The digital nature of the data allowed a complete 3D contextual visualization to be constructed whereby the individual sensory structures could be seen in relation to other inner ear structures. This included a detailed anatomy of the membranous labyrinth and nerve supply including the spatial configuration of the utricular and saccular maculae. This is a new way of undertaking temporal bone histology.