Abstract
Conventional microscopy has limitations in viewing the cochlear microstructures due to three-dimensional spiral structure and the overlying bone. But these issues can be overcome by imaging the cochlea in vitro with intravital multiphoton microscopy (MPM). By using near-infrared lasers for multiphoton excitation, intravital MPM can detect endogenous fluorescence and second harmonic generation of tissues. In this study, we used intravital MPM to visualize various cochlear microstructures without any staining and non-invasively analyze the volume changes of the scala media (SM) without removing the overlying cochlear bone. The intravital MPM images revealed various tissue types, ranging from thin membranes to dense bone, as well as the spiral ganglion beneath the cochlear bone. The two-dimensional, cross-sectional, and serial z-stack intravital MPM images also revealed the spatial dilation of the SM in the temporal bone of pendrin-deficient mice. These findings suggest that intravital MPM might serve as a new method for obtaining microanatomical information regarding the cochlea, similar to standard histopathological analyses in the animal study for the cochlea. Given the capability of intravital MPM for detecting an increase in the volume of the SM in pendrin-deficient mice, it might be a promising new tool for assessing the pathophysiology of hearing loss in the future.
Highlights
Despite its small size, the cochlea has a complex three-dimensional helical structure, consisting of the modiolus, a central pillar of spongy bone, and the spiral lamina winding around it
The organ of Corti consist of the tectorial membrane (TM), outer hair cells (OHC), inner hair cells (IHC), and supporting cells (Deiter cells, Hensen cells, and Claudius cells) (Figure 2D)
We describe a new method for microscopic analysis of the mammalian cochlea that offers several advantages over alternative histological preparations and allows for non-invasive spatial visualization of the scala media (SM) without removing the cochlear bone
Summary
The cochlea has a complex three-dimensional helical structure, consisting of the modiolus, a central pillar of spongy bone, and the spiral lamina winding around it. These structures include spaced filled with fluid [scala vestibuli, scala media (SM), and scala tympani] and the organ of Corti. The traditional microscopic methods used for examining these cochlear microstructures involve the creation of paraffin-embedded tissue sections. Many embedding sections may result in the loss of tissue antigenicity. Those make it difficult to conduct immunochemical studies on sections
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