Resistance exercise (RE) causes an acute increase of the muscle thickness (MT) considered to relate to an increase in tissue water content. Segmental bioelectrical impedance spectroscopy (S-BIS) is a tool used to noninvasively assess intra- and extra-cellular water (ICW and ECW, respectively) of a given limb segment. The purpose of the present study was to examine the change of MT, ultrasound echo intensity (EI), ICW, and ECW after 3 sets of exhaustive RE. Eighteen untrained young males (age, 25.4 ± 4.1 years) performed RE consisting of 3 sets of knee extension concentric and eccentric contractions with 80% of 1-repetition maximum to failure. The MT and EI of the quadriceps measured by ultrasonography, and ECW/ICW ratio of the thigh assessed by S-BIS before (baseline) and after each set of RE (PostEx1, 2, and 3). The changes (Δ) in MT, EI, and ECW/ICW ratio were calculated as values of PostEx minus baseline values. The values of MT, EI, and ECW/ICW ratio at PostEx3 were significantly higher than baseline (effect size: MT, 1.11; EI, 0.47; and ECW/ICW ratio, 0.45). In addition, ΔMT was significantly and moderately correlated with ΔECW/ICW ratio (r = 0.61). Integrated data showed weak but significant correlation between ΔEI and ΔECW/ICW ratio as well (r = 0.31). The present results suggest multiple sets of exhaustive knee extension RE induce the acute increase of EI and ECW/ICW ratio as well as MT. The acute increase of muscle size after exercise can be at least partly explained by relative ECW increase.