Appropriate temporal and spatial expression of osteopontin (OPN) in the female genital tract may be critical for successful embryo implantation and maintenance of gestation. Traditionally, experimental assessments of reproductive success have been limited to ex vivo dissection at a single time point to determine embryo number and size and are inadequate for ongoing study of the effect(s) of genetic manipulation on any individual gestation. To investigate the role of OPN in the maintenance of gestation, we developed a noninvasive, in vivo method of pregnancy surveillance suitable for murine application using magnetic resonance microscopy (MRM). Gravid wild-type mice (n =7) and mice with targeted disruption of one or both OPN alleles (OPN(-/-), n = 9; OPN(+/-), n = 3) underwent MRM on postcoital days 10.5, 15.5, and 19.5. Prenatal MRM images were used to determine embryo numbers and sizes. There were no significant differences in embryo numbers determined independently by two blinded observers (mean difference between observers = 0.04 embryos; p = 0.87). There was a significant effect of genotype on embryo size, with OPN(-/-) embryos significantly smaller at all gestational ages. However, targeted disruption of one or more OPN alleles had no effect on embryo number at any gestational age. Thus, MRM may be a powerful noninvasive method for in vivo prenatal developmental study of genetically engineered mice.