Objective: To develop a methodology for the measurement of balloon dilation (BD) effects on Eustachian Tube (ET) structure using Computerized Tomography (CT) images.Methods: The BD of the ET was performed on three cadaver heads (five ears) through the nasopharyngeal orifice. The axial CT images of the temporal bones were obtained before dilation, while an inflated balloon was in the lumen of ET, and after balloon removal in each ear. Utilizing Dicom images captured by the ImageJ software 3D volume viewer function, the anatomical landmark coordinates of the ET were matched with their pre- and post-dilation counterparts, and the longitudinal axis of the ET was captured with serial images. The histograms of the regions of interest (ROI) and three different lumen width and length measurements were obtained from captured images. The densities of air, tissue, and bone were determined with histograms as a baseline to determine the BD rate as a function of increased air in the lumen.Results: The small ROI box included the area of prominently dilated ET lumen after BD and best represented the visually obvious changes in the lumen, compared to the ROIs that extended the wider areas (longest and longer). Air density was the outcome measure for comparison with each corresponding baseline value. The average increase in air density in the small ROI was 64%, while the longest and long ROI boxes showed 44 and 56% increases, respectively.Conclusion:This study describes a method to image the ET and quantify the outcomes of BD of the ET using anatomical landmarks.