Although the synthesis of two-dimensional (2D) layered MoS2 nanomaterials have been developing rapidly, there are many technical issues in preparing MoS2 quantum dots (QDs) with photoluminescence property. Herein, we design a facile colloidal chemical route to prepare photoluminescent MoS2 QDs using the ammonium tetrathiomolybdate ((NH4)2MoS4) as precursors and oleyl amine as reducing agent. The optical property and structure of as-prepared MoS2 QDs are investigated systematically. Resultant MoS2 QDs exhibit fluorescence (λmax=575 nm; quantum yield, 4.4%), spherical morphology with uniform thickness of ~3 nm and excitation-dependent PL phenomenon. Moreover, resultant MoS2 QDs show size-dependent tunable photoluminescence in wide visible region. With the help of the amphiphilic compound, resultant MoS2 QDs could be transferred from organic to aqueous phase. MoS2 QDs in aqueous solution have many advantages, such as good dispersion, low toxicity and photoluminescent property which make them possess promising applications in optoelectronic and biological fields. In this study, the 293T cells are used as a model to evaluate the fluorescence imaging of MoS2 QDs. The results confirm fluorescent signal appears in cytoplasm which demonstrates asprepared MoS2 QDs could be used as a probe for real-time optical cellular imaging.