Recent studies show that the mitochondrial targeted antioxidant mitoquinone (MitoQ) reduces mitochondrial oxidative stress and prevents impaired mitochondrial dynamics. Focusing on the concept of mitochondrial reactive species as redox signaling molecules in insulin resistance rather than oxidative molecular damage as part of the ‘oxidative stress’, we wanted to examine the role of MitoQ, a mitochondrial antioxidant in the delivery of GLUT-4 translocation in skeletal muscle fibers. MitoQ is a shortened form of the antioxidant ubiquinol (a CoQ10) and is designed to be adsorbed to the inner mitochondrial membrane, as a potential therapeutic targeting mitochondrial. To study the hypothesis of improved GLUT4 translocation we used a genetically encoded biosensor GLUT4-7mycGFP plasmid in mouse skeletal FDB muscle. After the transfection (7 days) single FDB fibers were induced to insulin resistance by treating them with ceramides for twenty hours. After three-hour starvation insulin stimulation combined with MitoQ treatment was done to study GLUT4 movement in the membrane of the fat-induced fibers. Interestingly, microscopy image analysis showed that the clear increase in surface-exposed GLUT4 content induced by insulin were improved in the fibers treated with MitoQ compared to Control. In conclusion, MitoQ had positive effects related to GLUT4 translocation and improved glucose tolerance. Although, a lot of studies have suggested that mitochondria-targeted antioxidants may be worth considering as potentially helpful therapies more research needs to be done to recommend their use to patients with insulin resistance.