Abstract

BackgroundManual and semi-automatic analyses of images, acquired in vivo by confocal microscopy, are often used to determine the quality of corneal endothelium in the human eye. These procedures are highly time consuming. Here, we present two fully automatic methods to analyze and quantify corneal endothelium imaged by in vivo white light slit-scanning confocal microscopy.MethodsIn the first approach, endothelial cell density is estimated with the help of spatial frequency analysis. We evaluate published methods, and propose a new, parameter-free method. In the second approach, based on the stochastic watershed, cells are automatically segmented and the result is used to estimate cell density, polymegathism (cell size variability) and pleomorphism (cell shape variation). We show how to determine optimal values for the three parameters of this algorithm, and compare its results to a semi-automatic delineation by a trained observer.ResultsThe frequency analysis method proposed here is more precise than any published method. The segmentation method outperforms the fully automatic method in the NAVIS software (Nidek Technologies Srl, Padova, Italy), which significantly overestimates the number of cells for cell densities below approximately 1200 mm−2, as well as previously published methods.ConclusionsThe methods presented here provide a significant improvement over the state of the art, and make in vivo, automated assessment of corneal endothelium more accessible. The segmentation method proposed paves the way to many possible new morphometric parameters, which can quickly and precisely be determined from the segmented image.Electronic supplementary materialThe online version of this article (doi:10.1186/s12880-015-0054-3) contains supplementary material, which is available to authorized users.

Highlights

  • Manual and semi-automatic analyses of images, acquired in vivo by confocal microscopy, are often used to determine the quality of corneal endothelium in the human eye

  • Assuming that the cell density of the imaged section is representative for the whole endothelium, the endothelial cell density can be calculated with δf Segmentation we propose a new method to fully automatically segment the corneal endothelium

  • Density estimation We introduced the factor α to determine the endothelial cell density based on frequency analysis

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Summary

Introduction

Manual and semi-automatic analyses of images, acquired in vivo by confocal microscopy, are often used to determine the quality of corneal endothelium in the human eye. We present two fully automatic methods to analyze and quantify corneal endothelium imaged by in vivo white light slit-scanning confocal microscopy. Corneal endothelium The cornea is the frontal, transparent layer of the eye covering the pupil and iris. By maintaining the water fraction at 78%, the electrolyte balance ensures corneal transparency. Distortion of this equilibrium caused by damage of endothelial cells leads to swelling of the. Endothelial cell quantification is used as a screening method to elect patients for corneal refractive surgery and to determine the rate of endothelial cell loss after iris claw lens implantation

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