The application of vital dyes during vitreoretinal surgery greatly facilitates the visualization of different structures including the inner limiting membrane (ILM), epiretinal membranes or the vitreous. Indocyanine green was the first dye introduced for the staining of the ILM. Recent data indicate a potential toxic effect of ICG, with the underlying pathomechanisms not being understood completely. A novel dye for ILM staining is Brilliant blue G, which seems to have a much better safety profile compared to ICG. Another blue dye is Trypan blue to visualize epiretinal membranes. Trypan blue can either be injected into the airfilled globe or, after having been diluted with glucose into the fluid-filled globe. The presentation will focus on new safety studies on Brilliant blue obtained in animal studies and will discuss clinical data obtained in humans as well as ultrastructural features of tissue removed during surgery. In addition, other dyes, which are currently under investigation, such as Bromphenol blue, will be discussed and a perspective for future concepts will be given. The staining characteristics of these dyes when applied alone or in combination will be presented.