摘要： A few years ago, crucial graphene oxide (GO) features such as the carbon/oxygen ratio, number of layers, and lateral size were scarcely investigated and, thus, their impact on the overall optical biosensing performance was almost unknown. Nowadays valuable insights about these features are well documented in the literature, whereas others remain controversial. Moreover, most of the biosensing systems based on GO were amenable to operating as colloidal suspensions. Currently, the literature reports conceptually new approaches obviating the need of GO colloidal suspensions, enabling the integration of GO onto a solid phase and leading to their application in new biosensing devices. Furthermore, most GO-based biosensing devices exploit photoluminescent signals. However, further progress is also achieved in powerful label-free optical techniques exploiting GO in biosensing, particularly using optical fibers, surface plasmon resonance, and surface enhanced Raman scattering. Herein, a critical overview on these topics is offered, highlighting the key role of the physicochemical properties of GO. New challenges and opportunities in this exciting field are also highlighted.