In the past twenty years, the use of ultrasound-based methods has become a standard approach to measure tendon mechanical properties in vivo. Yet, the multitude of methodological approaches adopted by various research groups probably contributes to the large variability of reported values. The technique of obtaining and relating tendon deformation to tensile force in vivo has been applied differently, depending on practical constraints or scientific points of view. Divergence can be seen in i) methodological considerations such as the choice of anatomical features to scan and to track, force measurements or signal synchronisation and ii), in physiological considerations related to the viscoelastic behaviour or length measurements of tendons. Hence, the purpose of the present review is to assess and discuss the physiological and technical aspects connected to in vivo testing of tendon mechanical properties. In doing so, our aim is to provide the reader with a systematic, qualitative analysis of ultrasound-based techniques. Finally, a list of recommendations is proposed for a number of selected issues.