In both mammals and birds, the mechanical behavior of the middle ear structures is affected by the mechanical impedance of the inner ear. In this study, the aim was to quantify the acoustic impedance of the avian inner ear in the ostrich, which allows us to determine the effect on columellar vibrations and middle ear power flow in future studies. To determine the inner ear impedance, vibrations of the columella were measured for both the quasi-static and acoustic stimulus frequencies. In the frequency range of 0.3–4 kHz, we used electromagnetic stimulation of the ossicle and a laser Doppler vibrometer to measure the vibration response. At low frequencies, harmonic displacements were imposed on the columella using piezo stimulation and the resulting force response was measured with a force sensor. From these measurement data, the acoustic impedance of the inner ear could be determined. A simple RLC model in series of the impedance measurements resulted in a stiffness reactance of KIE = 0.20·1012 Pa/m3, an inertial impedance of MIE = 0.652·106 Pa s2/m3, and a resistance of RIE = 1.57·109 Pa s/m. We found that values of the inner ear impedance in the ostrich are one to two orders in magnitude smaller than what is found in mammal ears.