The condition of active muscles determines an individual′s ability to carry out daily activities and has implications for an athlete′s performance. Multi-frequency bioimpedance (mfBIA) is a non-invasive, well-known, validated, and much used method to assess muscle condition. However, it is rarely used to its full potential. Our aim was to apply mfBIA fully in the assessment of an adult healthy population, to compare muscle condition in different functional rested muscle groups, with age, and between men and women, and establish a control data set. Fifty healthy subjects (25 men/25 women) aged 20–69 years, participated. mfBIA measurements at a frequency range of 4–1000 kHz were taken from muscles of the lower and the upper extremities, the upper back, and the hand. Data were analyzed using ImpediMed software, giving Impedance, Resistance, Reactance, Phase Angle, Center Frequency, external and internal Resistance, and Membrane Capacitance. Differences between means were tested for statistical significance. A P value >0.05 was considered nonsignificant. While no difference in the mfBIA parameters was seen with age, a highly significant gender difference was seen. At rest, women′s muscles cf men's showed a significantly higher center frequency and intra- and extra-cellular resistance, while the membrane capacitance was lower. A set of values for mfBIA parameters for healthy adult individuals are given for some of the main muscles which are frequently part of muscle assessment. The documented gender difference in muscle condition at rest has important implications in work situations, during physical rehabilitation and when training for competitive sports.