Quantum dots (QDs) have the potential to be used in the multiplexed bioanalysis for their unique property:multi-color QDs can be excited at the same single wavelength light. In this work,high quan-tum yield multi-color core/shell QDs were prepared. After being water-solubilized by amphiphilic polymer based on self-assembling,the QDs would be labeled by two different IgGs and used in the multiplexed biodetection. Fluorescence resonance energy transfer (FRET) should be avoided in the multiplexed biodetection. In this work,spectral analysis showed that no FRET appeared in water-soluble QDs modified by amphiphilic polymer. The immunofluorescence in vitro indicated that the QDs-IgGs bioconjugates had excellent species-specific detection ability with nearly non-specific binding. The setting of this model will help to support the application of multi-color QDs in the multiplexed bioanalysis communities. Quantum dots (QDs) have the potential to be used in the multiplexed bioanalysis for their unique property: multi-color QDs can be excited at the same single wavelength light. In this work, high quan-tum yield multi-color core / shell QDs were prepared. After being water-solubilized by amphiphilic polymer based on self-assembling, the QDs would be labeled by two different IgGs and used in the multiplexed biodetection. Fluorescence resonance energy transfer (FRET) should be avoided in the multiplexed biodetection. work, spectral analysis showed that no FRET appeared in water-soluble QDs modified by amphiphilic polymer. The immunofluorescence in vitro indicated that the QDs-IgGs bioconjugates had excellent species-specific detection ability with nearly non-specific binding. The setting of this model will help to support the application of multi-color QDs in the multiplexed bioanalysis communities.