The twinkling sign (TS) observed in ultrasound imaging (e.g., color flow mode and pulse Doppler mode) has been reported in previous researches as a potential phenomenon to detect micro calcification in soft tissue. However, the mechanism of the twinkling sign has not been clearly understood yet. The authors investigated the effect of stiffness of surrounding tissue on the appearance of TS using the soft tissue-mimicking phantoms and a medical ultrasound device. The author used Poly (vinyl alcohol) hydro (PVA-H) gel as the material of phantom and developed three phantoms with different PVA concentration; 8 %wt, 10 %wt and 15 %wt those correspond to Young's modulus (E) as 50 kPa, 100 kPa and 230 kPa, respectively. Micro glass and CaCO3 particles were embedded in the phantoms as pseudo micro calcification. The authors observed TS in each phantom and analyzed the temporal average of TS. The temporal average of TS was largest in the 8 %wt (E = 50kPa) PVA-H gel phantom, and decreased with increasing the phantom stiffness. The result indicated that the micro oscillation of the particles had a close relationship with the occurrence of TS.