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Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow
Katerina Chatzimeletiou1, Alexandra Fleva2, Theodoros-Thomas Nikolopoulos1
1Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, 'Papageorgiou' General Hospital, Aristotle University Medical School, 56403 Thessaloniki, Greece.
Medicina (Kaunas, Lithuania)
|July 29, 2023
Summary
Both terminal-deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end labelling (TUNEL) assay and flow cytometry effectively detect sperm DNA fragmentation. These methods accurately assess male fertility by measuring DNA fragmentation index (DFI).
Area of Science:
- Reproductive biology
- Genetics
- Medical diagnostics
Background:
- Sperm DNA fragmentation involves breaks in sperm DNA strands, impacting male fertility.
- Common assessment methods include SCSA, SCD, SCGE-comet, and TUNEL assays.
- Evaluating sperm DNA fragmentation is crucial for male reproductive health assessments.
Purpose of the Study:
- To compare the efficiency and sensitivity of TUNEL assay via fluorescence microscopy and flow cytometry for analyzing sperm DNA fragmentation.
- To determine the reliability of these methods in assessing male fertility.
Main Methods:
- Semen samples were analyzed for standard characteristics using light microscopy.
- Sperm DNA fragmentation was assessed using both TUNEL via fluorescence microscopy and flow cytometry.
- Statistical analyses included Spearman's correlation and Mann-Whitney U test.
Main Results:
- No significant differences were found in the sperm DNA fragmentation index (DFI) between TUNEL and flow cytometry (p = 0.543).
- Significant negative correlations were observed between sperm motility and DFI (p < 0.01), and sperm concentration and DFI (p < 0.05).
- No significant DFI difference was noted between couples with repeated implantation failure and miscarriages (p = 0.352).
Conclusions:
- Both TUNEL via fluorescence microscopy and flow cytometry demonstrate high efficiency and sensitivity in detecting sperm DNA fragmentation.
- These methods are reliable tools for assessing male fertility and guiding reproductive health decisions.
