Aim:To determine whether glioma cells can be specifically and efficiently tar-geted by superparamagnetic iron oxide nanoparticle (SPIO)-fluoresceinisothiocyanate (FITC)-chlorotoxin (SPIOFC) that is detectable by magnetic reso-nance imaging (MRI) and optical imaging.Methods:SPIOFC was synthesized byconjugating SPIO with FITC and chlorotoxin.Glioma cells (human U251-MG andrat C6) were cultured with SPIOFC and SPIOF (SPIO-FITC),respectively.Neuralcells were treated with SPIOFC as the control for SPIOFC-targeted glioma cells.The internalization of SPIOFC by glioma cells was assessed by MRI and wasquantified using inductively-coupled plasma emission spectroscopy.The opticalimaging ability of SPIOFC was evaluated by confocal laser scanning microscopy.Results:Iron per cell of U251 (72.5±1.8 pg) and C6 (74.9±2.2 pg) cells cultured withSPIOFC were significantly more than those of U251 (6.6±1.0 pg) and C6 (7.1±0.8pg) cells incubated with SPIOF.The T_2 signal intensity of U251 and C6 cellscultured with SPIOFC (233.6±25.9 and 211.4±17.2,respectively) were substantiallylower than those of U251 and C6 cells incubated with SPIOF (2275.3±268.6 and2342.7±222.4,respectively).Moreover,there were significant differences in ironper cell and T_2 signal intensity between SPIOFC-treated neural cells (1.3±0.3;2533.6±199.2) and SPIOFC-treated glioma cells.SPIOFC internalized by gliomacells exhibited green fluorescence by confocal laser scanning microscopy.Conclusion:SPIOFC is suitable for the specific and efficient targeting of gliomacells.MRI and optical imaging in conjunction with SPIOFC can differentiateglioma cells from normal brain tissue cells. Aim: To determine whether glioma cells can be specifically and efficiently tar-geted by superparamagnetic iron oxide nanoparticle (SPIO) -fluoresceinisothiocyanate (FITC) -chlorotoxin (SPIOFC) that is detectable by magnetic reso- nance imaging (MRI) and optical imaging. Methods : SPIOFC was synthesized by conjugating SPIO with FITC and chlorotoxin. Glioma cells (human U251-MG andrat C6) were cultured with SPIOFC and SPIOF (SPIO-FITC), respectively. Neural cells were treated with SPIOFC as the control for SPIOFC-targeted glioma cells. The internalization of SPIOFC by glioma cells was assessed by MRI and wasquantified using inductively-coupled plasma emission spectroscopy. The optical imaging ability of SPIOFC was evaluated by confocal laser scanning microscopy. Results: Iron per cell of U251 (72.5 ± 1.8 pg) and C6 ( 74.9 ± 2.2 pg) cells cultured with SPIOFC were significantly more than those of U251 (6.6 ± 1.0 pg) and C6 (7.1 ± 0.8 pg) cells incubated with SPIOF. T_2 signal intensity of U251 and C6 cellscul (207.6 ± 25.9 and 211.4 ± 17.2, respectively) were substantiallylower than those of U251 and C6 cells incubated with SPIOF (2275.3 ± 268.6 and2342.7 ± 222.4, respectively). Moreover, there were significant differences in ironper cell and T_2 Signal intensity between SPIOFC-treated neural cells (1.3 ± 0.3; 2533.6 ± 199.2) and SPIOFC-treated glioma cells. SPIOFC internalized by gliomacells extracted green fluorescence by confocal laser scanning microscopy. Conlusion: SPIOFC is suitable for the specific and efficient targeting of glioma cells . MRI and optical imaging in conjunction with SPIOFC can differentiateglioma cells from normal brain tissue cells.