Grindstaff, TL and Threlkeld, AJ. Optimal stimulation parameters to detect deficits in quadriceps voluntary activation. J Strength Cond Res 28(2): 381–389, 2014—The purpose of this study was to determine optimal stimulation parameters and calculation methods to estimate quadriceps voluntary activation while minimizing participant discomfort. Twelve healthy adults (8 men and 4 women; mean ± SD, age = 36.8 ± 15.6 years, weight = 76.1 ± 12.9 kg, height = 170.2 ± 8.6 cm). Repeated maximal volitional isometric contractions (MVIC) were performed while imposing 4 stimulation combinations (10 or 2 pulses; 400 or 200 V; and variable or standardized current) with the quadriceps in a relaxed state (resting twitch [RT]) and during an MVIC. Quadriceps activation was quantified by calculating the central activation ratio and the percent activation. Discomfort was quantified using the visual analog scale. When comparing calculation methods between the same stimulation parameters, the central activation ratio calculation method produced quadriceps activation values that were significantly greater (p < 0.009) than those derived using the percent activation calculation method. The doublet pulse stimulus produced less discomfort during the RT (p < 0.04) and MVIC (p < 0.001) when compared with all other combinations using a train of stimuli (10 pulses). Correlations for all estimates of quadriceps activation were strong (r = 0.85–0.99, p < 0.001). A doublet pulse stimulus produced discomfort levels that were over 50% lower than a 10-pulse train of stimuli and correlated well (r > 0.88) with activation levels obtained with a 10-pulse train of stimuli. Therefore, the use of a doublet pulse stimulus provides quadriceps activation information equivalent to other methods while minimizing participant discomfort.