Mitochondria play a significant key in sperm metabolism as well as power generation. The mitochondria nucleolus is essential for several sperm activities. such as sperm capping, hyperactivation, and acrosome reactions, and oocyte fusion, mitochondrial membrane potential can be used to study sperm mitochondrial activity, the mitochondrial integrity of sperm is evaluated by examination of mitochondrial membrane potential MMP, which may be achieved by using fluorescent dyes like JC-1 probe is composed of 5,5,6,6-tetrachloro-1,1,3,3-tcarbocyanine iodide. To study the correlation between mitochondrial membrane potential Red MMP and sperm motility . This study included 42 male samples who were separated into two groups. (21 normozoospermic, 21 asthenozoospermia) through the period of attendance at the infertility clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University, and Um Al-Banin Center for Management and in vitro fertilization the collected semen was obtained and seminal fluid analysis was assisted according to (WHO,2021). fluorescent staining of the sample prepared is done by using an MMP Assay Kit (with JC-1) was performed. A positive significant correlation between sperm motility percentage and red MMP intensity (p < 0.001). Mitochondrial activity plays an essential function in sperm motility and male fertility Male infertility, Sperm Motility, Mitochondrial Membrane Potential, MMP Assay Kit (with JC-1).