Graphene, a unique 2D honeycomb structure consisting of conjugated sp2 hybridized carbon atoms with fascinating properties such as large theoretical surface area (2630 m2/g), better thermal conductivity (5000 W/m per K), special electronic conductivity (200,000 cm2/V per s), higher biocompatibility, low cost for scalable production, and ease of surface functionalization has been both conveniently and conventionally used to develop novel electronic materials including transparent conductors and ultrafast transistors. Recent understanding of various chemical properties of graphene however has facilitated its application range, which is now expanding beyond electronic and chemical applications toward biomedical areas, such as precise biosensing through graphene-quenched fluorescence, graphene-enhanced cell differentiation and growth, etc. Nanosized graphene derivatives are now finding regular use as in vivo/in vitro imaging agents and portable carriers for the targeted delivery of various anticancer drugs. Further research involving the functionalization and application of inorganic nanoparticles onto graphene has also gained considerable impetus. This chapter deals with a brief discussion on the historical backgrounds, physicochemical properties, synthetic methodologies, and potential use and application of graphene and its derivatives in the field of biomedicine, with special focus on the drug/gene delivery, imaging, therapeutic, and biosensing applications.