In this study, we aimed to develop new lipid based nanoparticles (LPNs) and radiolabeled LPNs with [99mTc]Tc-HMPAO to investigate its cell binding capacity comparatively with [99mTc]Tc-HMPAO on different cancer cells. According to obtained results, LPNs with zeta potential of -27.4 ± 0.95 mV, particle size of 93.5 ± 1.17 nm, and polydispersity index of 0.35 ± 0.04 were successfully developed. The optimum radiolabeling efficiency was found to be above 90% at 15-min of incubation time. The cell binding capacity of [99mTc]Tc-HMPAO-LPNs was found to be higher than [99mTc]Tc-HMPAO in cancer cell lines. The results demonstrated that [99m]Tc-HMPAO-LPNs may be a promising agent for cancer diagnosis alternatively to [99m]Tc-HMPAO. [ABSTRACT FROM AUTHOR]