We studied a slow- and a fast-twitch muscle fiber type of the perch that have different thin filament lengths. The force-sarcomere length relations were measured, and it was tested whether their descending limbs were predicted by the cross-bridge theory. To determine the predicted relations, filament lengths were measured by electron microscopy. Measurements were corrected for shrinkage with the use of I-band and H-zone periodicities. Thick filament lengths of the two fiber types were found to be similar (1.63 +/- 0.06 and 1.64 +/- 0.10 microns for slow- and fast-twitch fibers, respectively), whereas the thin filament lengths were clearly different: 1.24 +/- 0.10 microns (n = 86) for the slow-twitch type and 0.94 +/- 0.04 microns (n = 94) for the fast type. The descending limbs of the two fiber types are therefore predicted to be shifted along the sarcomere length axis by approximately 0.6 microns. Sarcomere length was measured on-line by laser diffraction in a single region in the center of the fibers. The passive force-sarcomere strain relation increased much more steeply in the slow-twitch fibers. The descending limb of the active force-sarcomere length relation of fast twitch fibers was linear (r = 0.92), but was found at sarcomere lengths approximately 0.1 micron greater than predicted. The descending limb of the slow-twitch fibers was also linear (r = 0.87) but was now found at sarcomere lengths approximately 0.05 microns less than predicted. The difference in position of the descending limbs of the two fiber types amounted to 0.5 microns, approximately 0.1 micron less than predicted. The difference between measured and predicted descending limbs was statistically insignificant.