The basilar membrane, an important functional part of the cochlea, is responsible for spectral separation of vibration signals transmitted from the stapes. In current study, scaled-up polymer membranes designed by mimicking the human basilar membrane were used for investigation of the frequency-separation characteristic. Displacement field formed on each polymer membrane was acquired by Laser Doppler scanning vibrometer and post-processed frequency-wise. The locations of the maximum displacement along the centerline were identified and collected for individual frequency range to produce the frequency-position map of individual polymer membrane. The influences of the membrane thickness and material properties on the variation of the frequency separability were discussed.