Description: Despite their widespread use in grazer–biofilm studies, stream exclusion cages have inherent physical properties that may alter benthic organism colonization and growth. We used laboratory studies and a field experiment to determine how exclusion cage design (size and material) alters light availability, water velocity, and benthic organism colonization. We measured light reduction by various plastic cage materials and flow boundary layer thickness across a range of exclusion cage sizes in the laboratory. We also deployed multiple exclusion cage designs based on commonly available materials into a second-order stream to assess algae and macroinvertebrate colonization differences among exclusion cages. All plastics reduced some light (190–700 nm wavelengths) and blocked more ultraviolet light than photosynthetically active radiation. Exclusion cage size did not influence flow boundary layer thickness, but larger exclusions tended to have higher velocity at the substrata surface. Despite light and water velocity differences, algal biomass, macroinvertebrate density, and community composition were similar between exclusion cage types. However, algal assemblages outside exclusion cages differed in composition and had higher biomass compared to inside exclusion cages. In terms of algal and macroinvertebrate colonization, plastic exclusion cage size and material appear to be flexible within the sizes tested, but differences can still exist between exclusion cage communities and those within the stream. Overall, artifacts of screened exclusion cages do not appear to introduce large bias in results of grazer–biofilm studies, but efforts to design exclusion cages that better mimic the natural system should continue.