Introduction Whole body high-frequency low-magnitude vibration (HFLMV) is routinely used in the medical field to improve bone quality in children. For this treatment, vibration is induced during a period when growth of the midpalatal suture is heavily influenced by the external environment. The midpalatal suture acts as a growth site responding to external signals stimulating deposition of bone on the sutural edges, facilitating transverse maxillary growth. Any additional mechanical strain such as HFLMV could modify the rate of bone remodelling at the interface of the two maxillary bones and affect natural growth. Objectives To evaluate the effects of HFLMV on the volume of the midpalatal suture (MPS) of rats. Materials and methods This study consisted of forty-two, five-week-old Fisher Strain male rats which were previously used in a study investigating the effects of vibration on hypofunctional teeth. The rats were randomly allocated into Vibration and Non-vibration groups. In the vibration groups, HFLMV was induced through whole body vibration platforms. The rats were kept in their cages and placed two at a time on the vibrating platform set at a magnitude of 0.3g and frequency of 30Hz. This stimulus was applied for 20 minutes per day, five days per week for a total of 30 days. The MPS was analysed as two separate volumes of interest in order to avoid the regions complicated by the pre-maxillomaxillary and palatomaxillary sutures. Three-dimensional micro-computed tomography (micro-CT) was used to quantify the volumes of the MPS. The measured volumes of the mid-palatal suture were adjusted by the length of the rat heads and statistical analysis was conducted. Results There was no statistically significant difference in the volumes of the MPS in any region between the Vibration and Non-vibration groups. Conclusion The findings of this 30-day animal study indicate that HFLMV applied through whole body vibration platform does not affect the volume of the MPS of rats.