Evaluation strategy for image acquisition protocols in confocal microscopy mosaic imaging of the cornea

In vivo confocal laser-scanning microscopy (CLSM) of the cornea provides image data from the corneal tissues with cellular resoultion. The ability to resolve the nerve fiber bundles of the sub-basal nerve plexus (SNP), in particular, opens the opportunity to use the SNP morphometry as a readily accessible, image-based quantitative biomarker for various neuropathic diseases. Several techniques have been proposed to overcome the small field of view of CLSM systems by capturing a dataset of many partially overlapping images covering an extended SNP area and creating a mosaic image from this dataset. So far, these mosaicking techniques have only been assessed qualitatively, by subjective visual inspection of the results. The present contribution describes an objective, quantitative evaluation strategy for these CLSM image montaging approaches by comparing the mosaicking results against ground truth data. Both the ground truth data and experimental datasets are acquired in a controlled experimental setup. Based on datasets from different specimens and different motion patterns, we benchmark our own mosaicking algorithm and report an "average of geometric distance" error value of ≤4.81 pixels. ... mehrWe provide the experimental data so that other researchers can evaluate and compare their own algorithms using the same base data.