We conducted clinical and experimental studies to investigate the effects of image-sharpening algorithms and color adjustments, which enabled real-time processing of live surgical images with a delay of 0.004 s. The images were processed with image-sharpening intensities of 0%, 12.5%, 25%, and 50% during cataract surgery, vitrectomy, peeling of epiretinal membrane, and peeling of internal limiting membrane (ILM) with the Ngenuity 3D visualization system. In addition, the images obtained with a yellow filter during the ILM peeling were processed with color adjustments. Five vitreoretinal surgeons scored the clarity of the images on a 10-point scale. The images of a 1951 United States Air Force grating target placed in no fluid (control), saline, and 0.1% and 1% milk solution were evaluated. The results showed that the mean visibility score increased significantly from 5.0 ± 0.6 at 0% to 6.4 ± 0.6 at 12.5%, 7.3 ± 0.7 at 25%, and 7.5 ± 0.9 at 50% (P < 0.001). The visibility scores during ILM peeling improved significantly with color adjustments (P = 0.005). In the experimental study, the contrast of the grating targets blurred by the 0.1% and 1% milk solution increased significantly by the image-sharpening procedure. We conclude that the image-sharpening algorithms and color adjustments improved the intraoperative visibility of 3D heads-up surgery.