Purpose <p>The aim of this study was to delineate and define, using ultrasound images, the boundaries of the different strata of the human tongue body, based on ultrasound examination of cadaveric samples.</p> Methods <p>The study was conducted using two fresh human cadavers from elderly male subjects, obtained through the Body Donation Program of the University of Chile. The tongues were dissected in sagittal and coronal planes, followed by detailed ultrasound examination using a Wisonic Piloter ultrasound equipment with a LH15-6 Hockey Stick linear probe. Metallic markers were used to demarcate the boundaries of the different strata during the ultrasound examination.</p> Results <p>Ultrasound images made it possible to distinguish between the mucosa and the underlying muscular strata of tongue. The mucosa appeared as three bands of different echogenicity, while the muscular strata exhibited varying echogenicities corresponding to their anatomical compositions. Stratum II was characterized by hypoechoic muscle fascicles with hyperechoic strands, while stratum III showed higher echogenicity with prominent hyperechoic speckles. The differentiation between stratum III and IV was clear in the central part of the tongue but less distinct laterally.</p> Conclusion <p>High-frequency ultrasound provided visualization of the detailed anatomy of the human tongue, including the delineation of its mucosal and muscular layers. This anatomical-ultrasonographic correlation could enhance the diagnostic accuracy of ultrasound for various pathological conditions affecting the tongue. Future studies should focus on evaluating variations in tongue anatomy based on age and gender to further refine ultrasound diagnostic capabilities.</p>