Sperm DNA integrity is an essential factor for successful fertilization and proper pregnancy progression. The terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) assay is commonly used for the estimation of the DNA fragmentation index. Analysis of TUNEL-stained sperm is often performed by flow cytometry, an approach that allows high-throughput analysis but in which any morphological information is lost. In this study, results of an automated image cytometry estimation of TUNEL-stained sperms were presented. The results of visual counting and automatic analysis were closely correlated, indicating that image cytometry is suitable for such analysis and may be applied in a clinical setting. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry