The survivin gene is highly expressed in pancreatic cancer. The purpose of this study was to design and synthesize functionalized magnetic iron oxide nanoparticles (MNPs) targeting survivin gene for the detection of pancreatic cancer. The pancreatic cancer cell line BxPC-3 with survivin gene expression was selected in this study. The healthy lung fibroblast cell was used as a control. Chitosan-coated MNPs (CS@MNPs) and antisense oligodeoxynucleotide of survivin gene were conjugated to MNPs to give Sur-MNPs. Fourier transform infrared spectroscopy was performed to confirm the conjunction of chitosan. The interactions of MNPs, CS@MNPs, and Sur-MNPs in BxPC-3 cells were observed, recorded and analyzed. The size, morphology, cell uptake, cytotoxicity and stability of those particles were assessed by transmission electron microscope, Prussian blue staining, MTT assay and agarose gel electrophoresis. The magnetic resonance signal intensities of pancreatic cells labeled with CS@MNPs and MNPs, and Sur-MNPs, were compared on T2-weighted images. The results demonstrated that the level of cellular uptake of CS@MNPs was higher than that of naked MNPs. The Sur-MNPs had a suitable size (12 nm sized core), high stability, no cytotoxicity and good water dispersion. Sur-MNPs did not accumulate in healthy lung fibroblast cells, while being taken up by BxPC-3 cells. The Sur-MNPs in BxPC-3 cells could be visualized on T2-weighted images, which suggested that Sur-MNPs could be used to detect the expression of survivin gene. Thus, Sur-MNPs may be a potential molecular imaging probe targeting survivin gene for early detection of pancreatic cancer cells. Copyright © 2012 John Wiley & Sons, Ltd.