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Background: The high prevalence of injuries in female athletes necessitates a course of action that not only enhances research in this field but also incorporates improved prevention programs and regular health monitoring of highly stressed structures such as tendons and muscles. Since myometry is already used by coaches and physiotherapists, it is important to investigate whether tissue stiffness varies in different types of sports, and whether such measures are affected by an acute training session. Methods: Myometric measurements of the Achilles tendon (AT) and soleus muscle (SM) were performed in the longitudinal plane and relaxed tendon position. In total, 38 healthy professional female athletes were examined, applying a quasi-experimental study design, with subgroup analysis performed for different sports. To investigate the stiffness of the AT and SM, 24 female handball and volleyball athletes performed a standardized maximal incremental performance test on a treadmill. In this subgroup, myometric measurements were taken before and after the exercise test. Results: The measurements showed no significant difference between the mean pre- (AT: 661.46 N/m; SM 441.48 N/m) and post-exercise stiffness (AT: 644.71 N/m; SM: 439.07 N/m). Subgroup analysis for different types of sports showed significantly lower AT and SM stiffness in swimming athletes compared to handball (p = 0.002), volleyball (p = 0.000) and hammer throw athletes (p = 0.008). Conclusions: Myometry can be performed on the same day as an acute training session in healthy female professional volleyball and handball athletes. Female swimmers have significantly lower AT and SM stiffness compared to female handball, volleyball and hammer throw athletes. These results show that the stiffness differences in the AT and SM can be assessed by myometry. [ABSTRACT FROM AUTHOR]