In the present work, a fluorescent gold nanoclusters (GNCs)/superparamagnetic (Fe3O4/GNCs) nanoprobe was prepared via a facile approach for the selective detection and imaging of human leukemica cancer cells (HL-60). (γ-Mercaptopropyl)trimethoxysilane (MPS) was used as a stabilizer to prepare functionalized GNCs. The prepared GNCs@MPS was then self-assembly decorated on the surface of Fe3O4@SiO2 nanoparticles followed by poly(ethylene glycol) dimethacrylate (PGD) addition at room temperature to form Fe3O4/GNCs nanoprobe. Surface functionalization of the Fe3O4/GNCs with the thiol-modified KH1C12 aptamer was done through thiol-en click reaction between PGD and the thiol group of the aptamer. An extensive characterization of the Fe3O4/GNCs revealed strong red fluorescence (λ em = 627 nm), T 2-based contrast agent for MRI and excellent colloidal and photo stability in buffer medium. So, the aptamer-functionalized Fe3O4/GNCs nanoprobe (Fe3O4/GNCs/Aptamer) is capable to uptake and dual-image HL-60 cancer cells from a mixture. Furthermore, the MRI signal intensity of the pictures decreased linearly with an increase in the concentrations of the nanoprobe. It is also enable to detect cancer cells from a range of concentrations 10 up to 200 cells μL-1. The fluorescent/magnetic characteristics of the nanoprobe are of great significance for MRI-based and fluorescence imaging and collection of HL-60 cancer cells which implies potential help for the development of early diagnosis of highly malignant human leukemia.