Abstract Acoustic microscopy can noninvasively measure acoustic properties, such as sound speed, density, and the attenuation coefficient of tissue sections or cells. The speed of sound can be estimated using a pulse-spectrum method; however, it may not be calculated accurately for samples with high attenuation. Moreover, a method for calculating the attenuation coefficient was proposed by Rohrbach et al., but the estimation error was large. Therefore, we propose a method for calculating the sou