Purpose Guided waves generated from bone cortex inevitably act on microbubbles flowing through skeletal muscle capillaries in contrast-enhanced ultrasound (CEUS) and might influence the image quality. However, the action mechanism underlying the guided waves influence is still unknown, especially under contrast pulse-inversion transmission mode. This study aimed to clarify the influence of guided waves on pulse-inversion CEUS, which was investigated via in vitro infusion experiments. Method Tibia guided waves were detected at pulse-inversion transmission and then characterized by using a short-time Fourier transform energy distribution. Using results at normal incidence as a baseline, the influence of guided wave dispersion on the contrast and resolution of pulse-inversion CEUS was investigated at an oblique incidence through continuous microbubbles infusion experiments in a vessel-tibia flow phantom. Results Frequency-dispersive property of tibia guided waves was observed at phases 0 degrees and 180 degrees, which improved the contrast of CEUS and reduced its resolution. Pulse-inversion CEUS balanced the contrast enhancement and resolution degeneration induced by guided waves. By contrast, contrast-to-tissue ratio of pulse-inversion CEUS increased by up to 109.1 +/- 13.2% (P < 0.05) due to guided waves and its resolution was up to 0.9 +/- 0.1 times that of baseline. Conclusions Alterations of contrast and resolution in pulse-inversion CEUS induced by guided waves might provide an additional assessment for the capillary perfusion in the skeletal muscle near the bone cortex.