An ultrasound indentation system for biomechanical assessment of soft tissues in vivo was developed. The pen-size, hand-held probe was composed of an ultrasound transducer and a load cell. The ultrasound transducer was at the tip of the probe serving also as the indentor. The thickness and deformation of the soft tissue layer were determined from the ultrasound echo. A compressive load cell was connected in series with the ultrasound transducer to record the force response. A validation experiment was performed on porcine tissues. Force and deformation acquired with the present system was in good comparison with those obtained from a Housfield material testing machine. Material constants were obtained via a curve-fitting procedure by predicting the force transient response from the deformation-time data using a quasilinear viscoelastic model. In addition, deformation in the fat and in the muscle could be differentiated. The potential applications of this type of indentation probes are many. The specific application of this current development is for stump tissue assessment in the design of prosthetics.