SUMMARY1. A technique has been developed with which the stiffness changes in frog skeletal muscle can be continuously recorded by measuring the propagation velocity of ultrasonic waves (3-7 MHz) with negligibly small perturbations to the contractile system.2. The resting muscle stiffness was 2 256+00002 x 109 N/m2 (S.D.) at 1-2°C (n = 10) and 2-480+0-007 x 109 N/M2 at 19-20°C (n = 12) in the longitudinal direction, and 2-223 + 04008 x 109 N/m2 at 1-2°C (n = 8) and 2 437 + 0 007 x 109 N/M2 at 19-20°C (n = 9) in the transverse direction.3. The resting muscle stiffness measured with ultrasonic waves was virtually insensitive to the resting force development, i.e. the extension of the parallel elastic component.4. The longitudinal muscle stiffness increased during isometric contraction at a rate faster than the force development. The amount of increase of the longitudinal stiffness in an isometric tetanus at 2 2 ,um sarcomere length was 24 ±+01 x 107 N/M2 at 1-2°C (n = 10) and 6-5 + 1-3 x 107 N/m2 at 19-20°C (n = 12).5. On the other hand, the transverse muscle stiffness decreased during isometric contraction at a rate faster than the force development. The amount of decrease of the transverse stiffness in an isometric tetanus at 22 jtm sarcomere length was 5-6+01 x 107 N/M2 at 1-2°C (n = 8) and 64+0-3 x 107 N/M2 at 19-20°C (n = 9).6. The amount of both the longitudinal and the transverse stiffness changes during an isometric tetanus decreased linearly with increasing sarcomere length, indicating that the stiffness changes during contraction reflect the formation of cross-links between the myofilaments.7. Both the longitudinal and the transverse stiffness increased when resting muscle was put into rigor state. The rigor muscle stiffness was insensitive to small stretches, i.e. the strain of the rigor cross-links.8. These results are discussed in connection with the behaviour of cross-bridges during isometric contraction and in rigor.