Carbon dots (CDs) as a new series of fluorescent nanomaterials have drawn great attention in recent years owning to their unique properties. In this paper, a simple carbonization approach to synthesize amino-functionalized CDs was developed by using chitosan as the carbon precursor. The as-prepared CDs possessed desirable amino function group on their surface and exhibited bright luminescence with absolute quantum yield (QY) of 4.34%, excitation-, pH-dependent and up-conversion fluorescence behaviors. Furthermore, we have investigated the cytotoxicity and biocompatibility of the as-prepared CDs, which demonstrated that the as-prepared CDs have the potential applications in biosensing, cellular imaging and drug delivery.