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Objectives: This research aims to apply an auditory display for tumor imaging using fluorescence data, discuss its feasibility for in vivo tumor evaluation, and check its potential for assisting enhanced cancer perception. Methods: Xenografted mice underwent fluorescence imaging after an injection of cy5.5-glucose. Spectral information from the raw data was parametrized to emphasize the near-infrared fluorescence information, and the resulting parameters were mapped to control a sound synthesis engine in order to provide the auditory display. Drag−click maneuvers using in-house data navigation software-generated sound from regions of interest (ROIs) in vivo. Results: Four different representations of the auditory display were acquired per ROI: (1) audio spectrum, (2) waveform, (3) numerical signal-to-noise ratio (SNR), and (4) sound itself. SNRs were compared for statistical analysis. Compared with the no-tumor area, the tumor area produced sounds with a heterogeneous spectrum and waveform, and featured a higher SNR as well (3.63 ± 8.41 vs. 0.42 ± 0.085, p < 0.05). Sound from the tumor was perceived by the naked ear as high-timbred and unpleasant. Conclusions: By accentuating the specific tumor spectrum, auditory display of fluorescence imaging data can generate sound which helps the listener to detect and discriminate small tumorous conditions in living animals. Despite some practical limitations, it can aid in the translation of fluorescent images by facilitating information transfer to the clinician in in vivo tumor imaging.