Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow Cytometry
<i>Background and Objectives</i>: 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 dispersi...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-07-01
|
| Series: | Medicina |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1648-9144/59/7/1313 |
| _version_ | 1827732496605773824 |
|---|---|
| author | Katerina Chatzimeletiou Alexandra Fleva Theodoros-Thomas Nikolopoulos Maria Markopoulou Glykeria Zervakakou Kyriakos Papanikolaou George Anifandis Anastasia Gianakou Grigoris Grimbizis |
| author_facet | Katerina Chatzimeletiou Alexandra Fleva Theodoros-Thomas Nikolopoulos Maria Markopoulou Glykeria Zervakakou Kyriakos Papanikolaou George Anifandis Anastasia Gianakou Grigoris Grimbizis |
| author_sort | Katerina Chatzimeletiou |
| collection | DOAJ |
| description | <i>Background and Objectives</i>: 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. <i>Materials and Methods</i>: 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. <i>Results:</i> 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 (<i>p</i> = 0.543). Spearman’s correlation analysis revealed a significant negative correlation between sperm motility (%) and sperm DNA fragmentation (<i>p</i> < 0.01), as well as between sperm concentration and sperm DNA fragmentation (<i>p</i> < 0.05). The Mann–Whitney U test showed no significant difference in the DFI among couples with repeated implantation failure (RIF) and miscarriages (<i>p</i> = 0.352). <i>Conclusions:</i> 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. |
| first_indexed | 2024-03-11T00:51:01Z |
| format | Article |
| id | doaj.art-9866e4a3142f4beb8382a3f218f69df2 |
| institution | Directory Open Access Journal |
| issn | 1010-660X 1648-9144 |
| language | English |
| last_indexed | 2024-03-11T00:51:01Z |
| publishDate | 2023-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Medicina |
| spelling | doaj.art-9866e4a3142f4beb8382a3f218f69df22023-11-18T20:24:59ZengMDPI AGMedicina1010-660X1648-91442023-07-01597131310.3390/medicina59071313Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow CytometryKaterina Chatzimeletiou0Alexandra Fleva1Theodoros-Thomas Nikolopoulos2Maria Markopoulou3Glykeria Zervakakou4Kyriakos Papanikolaou5George Anifandis6Anastasia Gianakou7Grigoris Grimbizis8Unit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, ‘Papageorgiou’ General Hospital, Aristotle University Medical School, 56403 Thessaloniki, GreeceDepartment of Immunology and Histocompatibility, ‘Papageorgiou’ General Hospital, 56403 Thessaloniki, GreeceUnit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, ‘Papageorgiou’ General Hospital, Aristotle University Medical School, 56403 Thessaloniki, GreeceDepartment of Immunology and Histocompatibility, ‘Papageorgiou’ General Hospital, 56403 Thessaloniki, GreeceFertilia by Genesis, IVF Unit, 54301 Thessaloniki, GreeceFertilia by Genesis, IVF Unit, 54301 Thessaloniki, GreeceDepartment of Obstetrics and Gynecology, School of Health Sciences, Faculty of Medicine, University of Thessaly, 41200 Larisa, GreeceDepartment of Immunology and Histocompatibility, ‘Papageorgiou’ General Hospital, 56403 Thessaloniki, GreeceUnit for Human Reproduction, 1st Department of Obstetrics & Gynaecology, ‘Papageorgiou’ General Hospital, Aristotle University Medical School, 56403 Thessaloniki, Greece<i>Background and Objectives</i>: 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. <i>Materials and Methods</i>: 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. <i>Results:</i> 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 (<i>p</i> = 0.543). Spearman’s correlation analysis revealed a significant negative correlation between sperm motility (%) and sperm DNA fragmentation (<i>p</i> < 0.01), as well as between sperm concentration and sperm DNA fragmentation (<i>p</i> < 0.05). The Mann–Whitney U test showed no significant difference in the DFI among couples with repeated implantation failure (RIF) and miscarriages (<i>p</i> = 0.352). <i>Conclusions:</i> 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.https://www.mdpi.com/1648-9144/59/7/1313spermDNA fragmentationtunel assayflow cytometryinfertility |
| spellingShingle | Katerina Chatzimeletiou Alexandra Fleva Theodoros-Thomas Nikolopoulos Maria Markopoulou Glykeria Zervakakou Kyriakos Papanikolaou George Anifandis Anastasia Gianakou Grigoris Grimbizis Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow Cytometry Medicina sperm DNA fragmentation tunel assay flow cytometry infertility |
| title | Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow Cytometry |
| title_full | Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow Cytometry |
| title_fullStr | Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow Cytometry |
| title_full_unstemmed | Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow Cytometry |
| title_short | Evaluation of Sperm DNA Fragmentation Using Two Different Methods: TUNEL via Fluorescence Microscopy, and Flow Cytometry |
| title_sort | evaluation of sperm dna fragmentation using two different methods tunel via fluorescence microscopy and flow cytometry |
| topic | sperm DNA fragmentation tunel assay flow cytometry infertility |
| url | https://www.mdpi.com/1648-9144/59/7/1313 |
| work_keys_str_mv | AT katerinachatzimeletiou evaluationofspermdnafragmentationusingtwodifferentmethodstunelviafluorescencemicroscopyandflowcytometry AT alexandrafleva evaluationofspermdnafragmentationusingtwodifferentmethodstunelviafluorescencemicroscopyandflowcytometry AT theodorosthomasnikolopoulos evaluationofspermdnafragmentationusingtwodifferentmethodstunelviafluorescencemicroscopyandflowcytometry AT mariamarkopoulou evaluationofspermdnafragmentationusingtwodifferentmethodstunelviafluorescencemicroscopyandflowcytometry AT glykeriazervakakou evaluationofspermdnafragmentationusingtwodifferentmethodstunelviafluorescencemicroscopyandflowcytometry AT kyriakospapanikolaou evaluationofspermdnafragmentationusingtwodifferentmethodstunelviafluorescencemicroscopyandflowcytometry AT georgeanifandis evaluationofspermdnafragmentationusingtwodifferentmethodstunelviafluorescencemicroscopyandflowcytometry AT anastasiagianakou evaluationofspermdnafragmentationusingtwodifferentmethodstunelviafluorescencemicroscopyandflowcytometry AT grigorisgrimbizis evaluationofspermdnafragmentationusingtwodifferentmethodstunelviafluorescencemicroscopyandflowcytometry |