Microplastics have been documented in aquatic and terrestrial ecosystems throughout the world. However, few studies have investigated microplastics in freshwater fish diets. In this study, water samples and three trophic levels of a freshwater food web were investigated for microplastic presence: amphipods (Gammarus lacustris), Yellowstone cutthroat trout (Oncorhynchus clarkii bouvieri), and lake trout (Salvelinus namaycush). Microplastics and other anthropogenic materials were documented in water samples, amphipods, and fish, then confirmed using FTIR (Fourier-transform infrared) and Raman spectroscopy. Our findings confirmed the presence of microplastics and other anthropogenic materials in three trophic levels of a freshwater food web in a high-elevation lake in a national park, which corroborates recent studies implicating the global distribution of microplastics. This study further illustrates the need for global action regarding the appropriate manufacturing, use, and disposal of plastics to minimize the effects of plastics on the environment.