Doppler strain rate imaging (SRI) was evaluated in vitro using a silicone strip phantom mimicking slowly moving tissue. A test apparatus was developed that enabled controlled strain experiments with variable strain and strain rate to be performed. SRI strain was measured at eight different calculated strains (range 5.7 to 63.4 %) at three different pump speeds with tissue velocity 0.1, 0.5 and 1.0 mm/s. The effect of varying tissue velocity and strain sample size on the measured SRI strain was elaborated. SRI strains agreed well with calculated values for strain when SRI strain was measured as the average over the whole strip cross-section and the strain sample size was 1.9 mm (mean difference = 2.78%, limits of agreement +/- 9.97% for tissue velocity 1.0 mm/s, n = 8). The variance was substantial if single central samples were used, especially for strain sample size of 0.8 mm (mean difference = -7.47%, limits of agreement +/- 20.90 for tissue velocity 0.5 mm/s, n = 24). Increasing the strain sample size to 1.9 mm removed some of the underestimation (giving mean difference of -4.46%, n = 24). We found low intra- and interobserver variation. This study indicates that, for the SRI method to give accurate estimates of strain, strain sample size should be in the region of 2 mm. Averaging over several ultrasound (US) beams increased the accuracy further.