Quantum dots (QDs) are very attractive for in vivo bio-imaging and therapeutic applications due to their relatively large absorption coefficient, high quantum yield, low level of photo bleaching, and large Stokes shift. However, two technical issues need to be resolved before they can be practically applied to in vivo bio-imaging applications: ensuring both reduced toxicity and efficient emission in the near-infrared (NIR) frequency range. Here we report a simple and reliable method to synthesize highly luminescent, NIRemitting Cu x In y S 2 /ZnS (CIS/ZnS) core-shell QDs for deep-tissue bio imaging applications. Offstoichiometric effects are utilized with 1-dodecanethiol as a reaction medium for thermolytic synthesis.The most important finding in our work is that at a high Cu/In ratio, the emission spectrum of CIS/ZnS QDs can be tuned to NIR frequencies with a high quantum yield up to approximately 65%. The maximum emission wavelengths of the synthesized QDs are 589 nm (QD 589 ) and 726 nm (QD 726 ) at a Cu/In ratio of 0.25 and of 1.8, respectively. Their feasibility for optical bio-imaging in a deep-tissue condition is investigated by the intramuscular injection of QD-loaded polymer microspheres in a mouse model. Our results show that more than 30% of the original emission of the QD 726 can be detected through biological tissue of 0.9 cm, whereas emission from the QD 589 is not detectable. Our investigation on the offstoichiometric effects of CIS QDs will contribute to the development of highly luminescent, NIR-emitting, cadmium-free QDs in the areas of tissue-level imaging, sensing, and therapeutics. † Electronic supplementary information (ESI) available: Size distribution histograms of the CIS QDs (Fig. S1); comparison of PLQY with a previous work (Fig. S2); the original curve of absorption spectra of the CIS QDs (Fig. S3); size distribution histograms of the CIS/ZnS QDs (Fig. S4); SEM images of the QD-loaded PMMA microspheres (Fig. S5). See