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Semen analysis constitutes the first biological step in the evaluation of male factor infertility (MFI) in an infertile couple. It is known that human spermatozoa generate reactive oxygen species in physiologic amounts, which play a role in sperm functions during sperm capacitation, acrosome reaction, and oocyte fusion. However, uncontrolled and excessive production of ROS, when it overwhelms the limited antioxidant defenses in semen, results in seminal oxidative stress. Recently, a substantial body of growing evidence suggests that such seminal oxidative stress is involved in many cases of idiopathic MFI. We have demonstrated that intracellular reactive oxygen species can be detected and measured in human spermatozoa by flow cytometry using the fluorescent probe 2',7'-dichlorofluorescein- diacetate (DCFH-DA). This method will help in assessing oxidative stress during basic semen analyses in assisted reproductive technology.
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