PurposeTo compare multi-slice (MS) MRI sequences of the pelvis acquired at rest and straining to dynamic midsagittal single-slice (SS) sequences for the assessment of pelvic organ prolapse (POP).MethodsThis IRB-approved prospective single-center feasibility study included 23 premenopausal symptomatic patients with POP and 22 asymptomatic nulliparous volunteers. MRI of the pelvis at rest and straining was performed with midsagittal SS and MS sequences. Straining effort, visibility of organs and POP grade were scored on both. Organ points (bladder, cervix, anorectum) were measured. Differences between SS and MS sequences were compared with Wilcoxon test.ResultsStraining effort was good in 84.4% on SS and in 64.4% on MS sequences (p= 0.003). Organ points were always visible on MS sequences, whereas the cervix was not fully visible in 31.1–33.3% on SS sequences. At rest, there were no statistically significant differences of organ point measurements between SS and MS sequences in symptomatic patients. At straining, positions of bladder, cervix, and anorectum were + 1.1 cm (± 1.8 cm), − 0.7 cm (± 2.9 cm), and + 0.7 cm (± 1.3 cm) on SS and + 0.4 mm (± 1.7 cm), − 1.4 cm (± 2.6 cm), and + 0.4 cm (± 1.3 cm) on MS sequences (p< 0.05). Only 2 cases of higher-grade POP were missed on MS sequences (both with poor straining effort).ConclusionMS sequences increase the visibility of organ points compared to SS sequences. Dynamic MS sequences can depict POP if images are acquired with sufficient straining effort. Further work is needed to optimize the depiction of the maximum straining effort with MS sequences.Graphical abstractSupplementary InformationThe online version contains supplementary material available at 10.1007/s00261-023-03944-8.