To determine the influence of spontaneous and instrumented vaginal delivery on objective measures of pelvic organ support.Prospective study at a university hospital with two study groups: vaginal spontaneous delivery (n = 26) and vacuum extraction (n = 49). Control group consisted of healthy nulliparous volunteers (n = 20). Participants underwent pelvic organ support evaluation by use of the pelvic organ prolapse quantification (POPQ) examination and postpartum functional cine magnetic resonance imaging (MRI).Significant differences for individual POPQ component measurements were noted for points Aa and Ba, TVL, and GH (spontaneous delivery versus control) and in addition for Ap, Bp, and D (vacuum extraction versus control). Significant differences for MRI measurements were observed for the position of bladder base, bladder neck, posterior fornix of the vagina, anorectal junction, hiatus perimeter and depth of rectocele.Considerable changes in pelvic organ support after vaginal delivery can be demonstrated by a reproducible and reliable clinical classification system (POPQ) and by functional cine magnetic resonance imaging.