Positively charged superparamagnetic iron oxide (SPIO) particles of maghemite were prepared in aqueous solution and subsequently stabilized with poly(ethylene oxide)-block-poly(glutamic acid) (PEO–PGA) at a hydrodynamic diameter of 60 nm. Depending on the amount of PEO–PGA used, this is accompanied by a switching of their zeta potentials from positive to negative charge (−33 mV). As a prerequisite for in vivo testing, the PEO–PGA coated maghemite nanoparticles were evaluated to be colloidally stable in water and in physiological salt solution for longer than six months as well in various buffer systems under physiological pH and salt conditions (AFM, dynamic light scattering). We excluded toxic effects of the PEO–PGA coated maghemite nanoparticles. We demonstrated by in vivo MR-imaging and ¹¹¹In measurements a biodistribution of the nanoparticles into the liver comparable to carboxydextran coated superparamagnetic iron oxide nanoparticles (Resovist^®) as a reference nanoscaled MRI contrast medium. This was enforced by a detailed visualization of our nanoparticles by electron microscopy of liver tissue sections. Furthermore, our results indicate that 15% of the injected PEO–PGA coated maghemite nanoparticles circulate in the blood compartment for at least 60 min after i.v. application.