A Novel Hybrid Software-Assisted Method to Evaluate Quantitatively Corneal Endothelium From Light Microscopy Images

Area of Science:

• Ophthalmology
• Biomedical Engineering
• Cell Biology

Background:

• Corneal endothelial cell density (ECD) and morphology are critical for eye banking and transplantation success.
• Current standard methods, light microscopy (LM) and specular microscopy (SM), have limitations in accuracy and evaluability.

Purpose of the Study:

• To develop and validate a novel Hybrid (HY) method integrating LM and SM for enhanced corneal endothelial cell analysis.
• To compare the performance of the HY method against traditional LM and SM techniques.

Main Methods:

• A Hybrid (HY) method was developed by processing LM images with SM-dedicated software for semi-automatic ECD, CV%, and HEX% determination.
• Bland-Altman analysis was used to assess agreement and inter-operator bias between LM, SM, and HY.
• Corneal evaluability was recorded across all three methods.

Main Results:

• The HY method demonstrated strong agreement with SM for ECD and HEX% determination, with clinically acceptable bias.
• HY showed significantly lower inter-operator variability in ECD and HEX% compared to LM.
• Corneal evaluability was significantly higher with HY (96.5%) and LM (96.5%) compared to SM (72.7%).

Conclusions:

• The HY method effectively enables quantitative analysis of corneal endothelial cells using LM images.
• HY provides a robust alternative, offering improved accuracy, reduced variability, and enhanced evaluability over traditional methods.