Objective: To investigate the feasibility and clinical significance of detecting sperm mitochondrial function by using Rh123/PI dual fluorescent staining and flow cytometry analysis, and to explore the relationship between the results of Rh123/PI dual fluorescent staining and seminal parameters.
Methods: Sixty-three semen samples were classified as normal (n=31) and abnormal (n=32) according to the World Health Organization guidelines. Rh123/PI dual fluorescent staining was then carried out to evaluate sperm mitochondrial function by flow cytometry analysis.
Results: Significant differences in Rh123+ PI-, Rh123- /PI+ and Rh123- /PI- were detected between the normal and abnormal semen samples (P < 0.05). There was a significant positive correlation between the Rh123+ PI- sperm and sperm motility and a significant inverse correlation between Rh123+ PI- and immotile sperm. But the Rh123- PI+ sperm showed a contrary relationship with Rh123+ PI-. A significant inverse correlation was also observed between the Rhl23- /PI- sperm and sperm concentration in the abnormal group.
Conclusion: Rh123/PI dual fluorescent staining and flow cytometry analysis can readily and quickly detect sperm mitochondrial function and be used to evaluate semen quality.