Muscle injuries in the lower limb are common among professional football players. Classification is made according to severity and is diagnosed with radiological assessment as: grade I (minor strain or minor injury), grade II (partial rupture, moderate injury) and grade III (complete rupture, severe injury). Tetrapolar localized bioimpedance analysis (BIA) at 50 kHz made with a phase-sensitive analyzer was used to assess damage to the integrity of muscle structures and the fluid accumulation 24 h after injury in 21 injuries in the quadriceps, hamstring and calf, and was diagnosed with magnetic resonance imaging (MRI). The aim of this study was to identify the pattern of change in BIA variables as indicators of fluid [resistance (R)] and cell structure integrity [reactance (Xc) and phase angle (PA)] according to the severity of the MRI-defined injury. The % difference compared to the non-injured contralateral muscle also measured 24-h after injury of R, Xc and PA were respectively: grade I (n = 11; -10.4, -17.5 and -9.0%), grade II (n = 8; -18.4, -32.9 and -16.6%) and grade III (n = 2; -14.1, -52.9 and -43.1%), showing a greater significant decrease in Xc (p < 0.001). The greatest relative changes were in grade III injuries. However, decreases in R, that indicate fluid distribution, were not proportional to the severity of the injury. Disruption of the muscle structure, demonstrated by the localized determination of Xc, increased with the severity of muscle injury. The most significant changes 24 h after injury was the sizeable decrease in Xc that indicates a pattern of disrupted soft tissue structure, proportional to the severity of the injury.