Recent advantages in the imaging of human corneal endothelial cells

Abstract

Sixty years after the understanding of their crucial role in corneal transparency maintenance, endothelial cells (ECs) are back under the spotlight since the development of endothelial keratoplasty and since new drugs and bioengineering techniques are in the pipeline. Accurate and informative imaging techniques are necessary to objectively assess the efficacy of each new therapeutic option. We will describe 3 recent advances in this field. 1/a simple laboratory assay coupled with image analysis that facilitates the assessment of toxicity of any kind of process liable to interfere with ECs. 2/a software that uses optimized algorithms of 3D reconstruction and of cell segmentation in order to increase the accuracy of EC count in eye banks by avoiding parallax errors that occur when ECs are counted in the deep posterior folds of stored corneas. 3/a 3D mapping of the subcellular localization of a set of proteins characteristic of ECs (in absence of a unique marker). We used confocal imaging of fluorescence immunolabelling of structural and functional proteins of normal human ECs. This 3D map could prove useful to characterize cells obtained during bioengineering processes that are candidate to become advanced‐therapy medicinal products.