Voluntary activation levels during lengthening, isometric, and shortening contractions (angular velocity 60°/s) were investigated by using electrical stimulation of the femoral nerve (triplet, 300 Hz) superimposed on maximal efforts. Recruitment of fiber populations was investigated by using the phosphocreatine-to-creatine ratio (PCr/Cr) of single characterized muscle fibers obtained from needle biopsies at rest and immediately after a series of 10 lengthening, isometric, and shortening contractions (1 s on/1 s off). Maximal voluntary torque was significantly higher during lengthening (270 ± 55 N·m) compared with shortening contractions (199 ± 47 N·m, P < 0.05) but was not different from isometric contractions (252 ± 47 N·m). Isometric torque was higher than torque during shortening (P < 0.05). Voluntary activation level during maximal attempted lengthening contractions (79 ± 8%) was significantly lower compared with isometric (93 ± 5%) and shortening contractions (92 ± 3%, P < 0.05). Mean PCr/Cr values of all fibers from all subjects at rest were 2.5 ± 0.6, 2.0 ± 0.7, and 2.0 ± 0.7, respectively, for type I, IIa, and Ilax fibers. After 10 contractions, the mean PCr/Cr values for grouped fiber populations (regardless of fiber type) were all significantly different from rest (1.3 ± 0.2, 0.7 ± 0.3, and 0.8 ± 0.6 for lengthening, isometric, and shortening contractions, respectively; P < 0.05). The cumulative distributions of individual fiber populations after either contraction mode were significantly different from rest (P < 0.05). Curves after lengthening contractions were less shifted compared with curves from isometric and shortening contractions (P < 0.05), with a smaller shift for the type Ilax compared with type I fibers in the lengthening contractions. The results indicate a reduced voluntary drive during lengthening contractions. PCr/Cr values of single fibers indicated a hierarchical order of recruitment of all fiber populations during maximal attempted lengthening contractions.