OL volume histograms from all the samples showing the OL volume distribution from 20 to 1200 μm³.

OL volume average and standard deviations are provided in Table 2. Note that the unusual distribution of T2 may be due to the poor resolution of the scan (see Methods).</p

The OL concentration using the two segmentation approaches.

Dark blue represents the MT segmentation with volumes from 10–2000μm3, light blue is the same segmentation method but with volumes from 20–2000 μm3, and grey represents the DLM segmentation for OL volumes between 10–2000μm3.</p

Numbers of OL from all different samples.

Numbers of OL from all different samples.</p

Light-sheet fluorescent microscopy images of a cleared pig petrous bone.

All images are from the same Z-depth with (green) autofluorescence and (red) DRAQ5 signals. Low magnification images show the cleared bone tissue from the cochlear cavities towards the outer bone surface. High magnification images of areas indicated by the rectangles are shown. The local concentrations of DRAQ5 stained nuclei can best be seen in the high magnification image.</p

2D representation of the temporal bone including a longitudinal cross section of the otic capsule.

(A) A slice from a low-resolution μ-CT tomogram obtained from the pig right temporal bone before extraction of the petrous bone showing the otic capsule in the grey rectangle. (B) SEM backscattered electron automated image map of a transverse section of a pig petrous bone after extraction, sectioning and polishing. The mastoid bone samples were collected from the same location (T) from two different pigs (Fig 2A). The locations of petrous bone samples from the outer and inner layers (OP and IP) are shown in Fig 2B. These samples were collected from the same locations in two different pigs.</p

Morphometric analysis of lacunae from all samples.

Morphometric analysis of lacunae from all samples.</p

Flow chart of the pig samples in this study.

The animal used is indicated at the top row, and the samples extracted from each animal are indicated in the second row. The third row shows sample preparation and then the flow of analyses after each experiment.</p

Fig 3 -

High resolution μ-CT tomogram slice of the inner layer sample (IP2) after MT (top) and DLM (bottom) segmentation. Left images show the whole field of view at the same slice location, and right images show insets of the OL labels as obtained from the two different segmentation approaches. Colors are a qualitative indication of the volume of the OL labels. Scale bars = 100μm.</p

S1 File -

(DOCX)</p