The temperature dependence of the dielectric parameters of tissues and tissue-mimicking phantoms is very important for non-invasive temperature measurement in medical applications using microwaves. We performed measurements of this dependence in the temperature range of 25–50°C using distilled water as a reference liquid commonly used in dielectric property studies. The results were compared with the literature model in the frequency range of 150–3000 MHz. Using this method, the temperature dependence of dielectric parameters of a new muscle tissue-mimicking phantom based on agar, polyethylene powder, and polysaccharide material TX-151 was measured in the temperature range of 25–50°C. The temperature dependence of the dielectric properties of this new muscle phantom was fitted to that of the two-pole Cole–Cole model and the deviation of the results between measured and modeled data was quantified.