In the human eye, accommodation is essential for functional vision. However, the mechanisms regulating accommodation and the ocular parameters affecting aberrations remain to be explored. In order to measure the alterations of ocular aberration and crystalline lens biometry during dynamic accommodative stimuli, we designed an optical coherence tomography with ultra-long penetration depth (UL-OCT) combined with a Shack–Hartmann wavefront sensor (SHWFS). This integrated set up measures human eye's anterior segment as well as monochromatic high-order aberrations (HOAs) with 6 μm resolution and (1/20) λ accuracy. A total of 10 healthy volunteers without ocular diseases were examined. Upon exposure to accommodative stimuli, the wavefront aberrations became larger. Among the anterior segment biometry, the anterior crystalline lens demonstrated significant curvature during accommodation and was the major cause of high-order aberration. These findings suggest that the front surface of the crystalline lens can significantly affect variation among aberrations, which is a key factor underlying the quality of human vision.