Trypan blue (TB) is a blue vital dye with fine color properties to stain the anterior lens capsule and thereby may facilitate capsulorrhexis during cataract surgery. In addition, the blue stain may assist in the visualization of various preretinal membranes and tissues during vitreoretinal surgery in a procedure also called chromovitrectomy. TB has demonstrated great binding affinity for the glial epiretinal membranes, although it remains yet to be determined in which circumstances the dye may color the vitreous and internal limiting membrane. Most studies suggest that 0.06% TB does not pose harm to the retina, but at higher concentrations further investigation is necessary. In this paper, various aspects of the application of TB for chromovitrectomy are discussed including laboratory investigations, surgical technique and clinical outcomes.