Muscle biopsy samples were obtained from healthy subjects in order to evaluate quantitative differences in single fibres of substrate (glycogen and triglyceride) and ion concentrations (Na+ and K+) as well as enzyme activity levels (succinate-dehydrogenase, SDH; phosphofructokinase, PFK; 3-hydroxyacyl-CoA-dehydrogenase, HAD; myosin ATPase) between human skeletal muscle fibre types. After freeze drying of the muscle specimen fragments of single fibres were dissected out and stained for myofibrillar-ATPase with preincubations at pH's of 10.3, 4.6, and 4.35. Type I (“red”) and II A, B, and C (“white”) fibres could then be identified. Glycogen content was the same in different fibres, whereas triglyceride content was highest in Type I fibres (2–3 × Type II). No significant differences were observed for Na+ and K+ between fibre types. The activity for the enzymes studied were quite different in the fibre types (SDH and HAD, Type I ≊ 1.5 × Type II; PFK Type I ≊ 0.5 × Type II; Myosin ATPase Type I ≊ 0.4 × Type II). The subgroups of Type II fibres were distinguished by differences in both SDH and PFK activities (SDH, Type II C > A > B; PFK, Type II B > A ≊ C). It is concluded that contractile and metabolic characteristics of human skeletal fibres are very similar to many other species. One difference, however, appears to be that no Type II fibres have an oxidative potential higher than Type I fibres.