Background/Objectives To evaluate biometric changes throughout the anterior chamber during accommodation and presbyopia using single image acquisition swept-source anterior-segment optical coherence tomography (AS-OCT). Subject/Methods Anterior-segment images were obtained using a new swept-source AS-OCT device (ANTERION, Heidelberg Engineering) from healthy volunteers (n = 71) across two centers in this prospective observational case series. In one image acquisition, cornea through posterior lens, including the ciliary muscle on both sides of the right eye, was imaged. Subjects undertook no accommodative effort and −1, −3, and −5 D of target vergence. Two-way repeated measures ANOVA modeling was performed for ciliary muscle measurements, lens parameters, aqueous depth (AD), and pupil diameter (PD). The first ANOVA factor was accommodative stimuli, and the second factor included age and refractive status.Results Maximum ciliary muscle thickness increased with accommodative stimuli (p < 0.001), while the distance from the scleral spur to the maximal point on the ciliary muscle and posterior ciliary muscle thickness (CMT2) decreased (p < 0.001-0.002). Older individuals showed no accommodative changes for ciliary muscle parameters, lens thickness, lens vault, PD, and AD (p = 0.07-0.32). Younger-and middle-aged eyes showed statistically significant accommodative structural alterations for these endpoints (p < 0.001-0.002), but with different patterns, including early loss of CMT2 contraction in middle-aged eyes. Within the middle-aged group, myopic eyes maintained better capacity for accommodative structural change. Conclusions Swept-source AS-OCT demonstrated multiple simultaneous anterior-segment biometric alterations in single acquisition images, including early loss of posterior ciliary muscle function and better maintained capacity for anteriorsegment structural change in myopia.