Objective To investigate the effect of super paramagnetic iron oxide nanoparticles (SPIO) on the function of contrast agent by Ultrasonic(US) and Magnetic Resonance Imaging (MRI). Methods The perluorocarbon-filled albumin-SPIO microbubbles were synthesis with an ultrasonic processor and used as US/MRI multifunctional contrast agents. The physical characterizations were accessed by transmission electron microscopy (TEM), X-ray energy dispersive spectroscopy, and UV-visible light absorption spectrum. The influence of SPIO concentration on the function of contrast agents was further studied by US/MRI. Results The ultrasound echo response will be enhanced with the SPIO concentration increasing. The enchancement of r2 value might be attributed to the synergetic magnetism effect arising from the aggregation and stabilization of multiple SPIOs in microbubble shells. The multifunctional microbubbles may be superior negative MRI contrast agents beyond regular SPIOs. Conclusion The contrast agents could improve the US/MRI resolution and contrast, which is helpful to the design method of the multifunctional microbubble contrast agent.