Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow Cytometry
Katerina Chatzimeletiou,
Alexandra Fleva,
Theodoros-Thomas Nikolopoulos,
Maria Markopoulou,
Glykeria Zervakakou,
Kyriakos Papanikolaou,
George Anifandis,
Anastasia Gianakou,
Grigoris Grimbizis
Affiliations
Katerina Chatzimeletiou
Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, ‘Papageorgiou’ General Hospital, Aristotle University Medical School, 56403 Thessaloniki, Greece
Alexandra Fleva
Department of Immunology and Histocompatibility, ‘Papageorgiou’ General Hospital, 56403 Thessaloniki, Greece
Theodoros-Thomas Nikolopoulos
Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, ‘Papageorgiou’ General Hospital, Aristotle University Medical School, 56403 Thessaloniki, Greece
Maria Markopoulou
Department of Immunology and Histocompatibility, ‘Papageorgiou’ General Hospital, 56403 Thessaloniki, Greece
Glykeria Zervakakou
Fertilia by Genesis, IVF Unit, 54301 Thessaloniki, Greece
Kyriakos Papanikolaou
Fertilia by Genesis, IVF Unit, 54301 Thessaloniki, Greece
George Anifandis
Department of Obstetrics and Gynecology, School of Health Sciences, Faculty of Medicine, University of Thessaly, 41200 Larisa, Greece
Anastasia Gianakou
Department of Immunology and Histocompatibility, ‘Papageorgiou’ General Hospital, 56403 Thessaloniki, Greece
Grigoris Grimbizis
Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, ‘Papageorgiou’ General Hospital, Aristotle University Medical School, 56403 Thessaloniki, Greece
Background and Objectives: Sperm DNA fragmentation refers to any break in one or both of the strands of DNA in the head of a sperm. The most widely used methodologies for assessing sperm DNA fragmentation are the sperm chromatin structure assay (SCSA), the sperm chromatin dispersion assay (SCD), the single-cell gel electrophoresis assay (SCGE–comet), and the terminal-deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end labelling (TUNEL) assay. The aim of this study was to compare the efficiency and sensitivity of the analysis of sperm DNA fragmentation using TUNEL via fluorescence microscopy, and flow cytometry. Materials and Methods: Semen samples were collected and analyzed for standard characteristics using light microscopy, and for sperm DNA fragmentation using both TUNEL via fluorescence microscopy, and flow cytometry. Results: There were no significant differences in the values of the sperm DNA fragmentation index (DFI) obtained when the analysis was performed using TUNEL or flow cytometry (p = 0.543). Spearman’s correlation analysis revealed a significant negative correlation between sperm motility (%) and sperm DNA fragmentation (p p p = 0.352). Conclusions: Both methods (TUNEL via fluorescence microscopy, and flow cytometry) have a high efficiency and sensitivity in accurately detecting sperm DNA fragmentation, and can be effectively used to assess male fertility.
