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Updated: May 9, 2026
Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects
Published on: April 21, 2022
Evaluation of conventional semen parameters, intracellular reactive oxygen species, DNA fragmentation and dysfunction
Leila Rashki Ghaleno1, Mojtaba Rezazadeh Valojerdi, Ehsan Janzamin
1Department of Embryology at Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, P. O. 19395-4644, Tehran, Iran, lrashki@yahoo.com.
Purpose:
The aim of this study was to evaluate the conventional sperm parameters, level of intracellular reactive oxygen species (ROS), DNA fragmentation (DF) and dysfunction of mitochondrial membrane potential (MMP) after semen preparation techniques with flow cytometry.
Methods:
Semen samples were obtained from 28 men with normal semen analysis according to WHO (world health organization). Each was divided into three equal parts for processing with routine techniques: conventional swim up (CSW), direct swim up (DSW) and density gradient centrifugation (DGC). The conventional sperm parameters were evaluated with computer-assisted sperm analyzer (CASA) and the level of intracellular ROS, dysfunction of MMP and DF were determined with flow cytometry procedure.
Results:
Conventional sperm parameters such as motility, progressive motility and normal morphology increased after sperm processing by CSW and DGC compared to DSW. A significant increase in intracellular H₂O₂ (p < 0.05) was demonstrated in the CSW versus DSW technique, while processed sperm by the DSW procedure showed a significant increase in the percentage of dysfunction of MMP and intracellular O₂(•-) (p < 0.05) when compared with CSW and DGC techniques. Additionally, a high mean of DF (p < 0.05) was observed in the DGC technique as compared to CSW.
Conclusion:
Data from flow cytometry study demonstrated that intracellular H₂O₂ and DF increased after CSW and DGC processing techniques, respectively, whereas the level of intracellular O₂(•-) and dysfunction of MMP only increased after the DSW processing technique.
