Objective: To study single sperm boar motility using electrical impedance measurements in a microfluidic system. Design: Comparison of the optical data and electrical impedance data. Setting: Research laboratory at a university. Animal(s): Boar semen sample were used. Intervention(s): A microfluidic system is developed that is able to spatially confine single boar sperm cells and allows noninvasive analysis of their motility on the single cell level. Using this system, the single sperm motility was affected by changing the temperature or adding chemical stimuli (caffeine). The retrieved electrical impedance and video data were processed using Matlab. Main Outcome Measure(s): The sperm beat frequency and amplitude determined from the electrical impedance and video data. Result(s): The electrically measured sperm beat frequency was verified by optical analysis and in correspondence. Furthermore the microfluidic platform allowed single sperm analysis by altering the sperm by temperature and chemical stimuli. Conclusion(s): This platform could be exploited as a potential tool to study sperm cells on the single cell level and to perform advanced sperm selection for intracytoplasmic sperm injection (ICSI) applications. (C) 2016 by American Society for Reproductive Medicine.