Insect auditory transduction occurs within a scolopidium—a multicellular unit comprised of one or two auditory neurons enclosed in a supporting cell called the scolopale cell. In a tympanic ear, the apical end of the scolopidium is embedded in a cap cell that connects the scolopidium in series to the tympanum. The mechanism by which vibrations at the tympanum elicit the transducing current in the scolopidium remains highly speculative due to the absence of direct observation of sound-evoked motion in the scolopidium. In this study, we examined scolopidia from auditory neurons in ex-vivo preparations of Müller’s organ obtained from the desert locust, Schistocerca gregaria. We used Optical Coherence Tomography (OCT) to characterize background motions of Müller’s organ in the region where the scolopidia is located. Subsequently, we employed a CMOS-based high-photon efficiency (capturing 95% of photons), high-speed camera to characterize the relative motion within the scolopidium at about ten thousand frames per second in response to sound stimulation from 1 to 5 kHz. The characterization of the relative motion within the scolopidium was achieved by subtracting the background motion of Müller’s organ using image processing techniques. The obtained characterization reveals the complex motions of Müller’s organ itself and that of the scolopidium, implicating the role of these motions in locust auditory transduction. Additionally, we propose a possible gating mechanism of insect auditory transduction based on these observations.