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The use of magnetic nanoparticles (MNPs) in biomedical applications requires the quantitative knowledge of their quantitative distribution within the body. AC Biosusceptometry (ACB) is a biomagnetic technique recently employed to detect MNPs in vivo by measuring the MNPs response when exposed to an alternate magnetic field. The ACB technique presents some interesting characteristics: non-invasiveness, low operational cost, high portability, and no need for magnetic shielding. ACB conventional methods until now provided only qualitative information about the MNPs' mapping in small animals. We present a theoretical model and experimentally demonstrate the feasibility of ACB reconstructing 2D quantitative images of MNPs' distributions. We employed an ACB single-channel scanning approach, measuring at 361 sensor positions, to reconstruct MNPs' spatial distributions. For this, we established a discrete forward problem and solved the ACB system's inverse problem. Thus, we were able to determine the positions and quantities of MNPs in a field of view of 5 × 5 × 1   cm 3 with good precision and accuracy. The results show the ACB system's capabilities to reconstruct the quantitative spatial distribution of MNPs with a spatial resolution better than 1   cm , and a sensitivity of 1.17   mg of MNPs fixed in gypsum. These results show the system's potential for biomedical application of MNPs in several studies, for example, electrochemical-functionalized MNPs for cancer cell targeting, quantitative sensing, and possibly in vivo imaging. [ABSTRACT FROM AUTHOR]