Isometric peak twitch force (stimulation frequency 0.5/s; 29.5-30.5 degrees C) was correlated with sarcomere length in isolated papillary muscles of the rabbit. Sarcomere length was measured from photographic recordings (1.5 ms exposure time) performed at rest between contractions and at the time of isometric peak twitch force. The sarcomere length at rest was found to be relatively uniform throughout the preparation and to be linearly related to the overall muscle length within the range Lmax-0.85Lmax. The distribution of sarcomere lengths increased considerably as the muscle went from rest to activity. Studies of surface markers showed different degrees of shortening (or elongation) of individual segments along the length of the preparation. The mean resting sarcomere length at Lmax (the optimum muscle length for force production) was 2.44 +/- 0.01 micron (grand mean +/- S.E., 7 muscles). The means active sarcomere length at Lmax was 2.29 +/- 0.04 micron. Active force declined steeply as the muscle length was reduced below Lmax. At a resting sarcomere length of 2.0 micron, active force was approximately 1/3 of the maximum. The observed differences between the length-tension relat-onships in myocardium (twitch responses) and skeletal muscle (tetanic contractions) are discussed on the basis of a length dependency of the activation process in cardiac muscle.