Despite the rise of marine diatoms in the world's oceans throughout the Cretaceous, only a handful of fossil localities worldwide detail invasion of freshwater habitats by diatoms commencing in the Late Cretaceous. We report on the occurrence of numerous freshwater diatom specimens and species from the Battle Formation, an extensive freshwater locality in western Canada that formed in the Late Cretaceous approximately 66.5 million years ago (Ma). The formation represents one of the oldest known localities worldwide harboring definitive remains of freshwater diatoms, contains the oldest known freshwater specimens of the centric diatom genus Aulacoseira, and confirms that these early Aulacoseira colonizers formed filaments linked together with interdigitating spines. We further document a high diversity of araphid pennate diatoms belonging to the order Fragilariales. Seven pennate morphotypes were uncovered, six of which definitively lacked a raphe and whose closest modern relatives are in the genera Fragilariforma, Fragilaria, and Stauroforma. Given the extensive coverage of the Battle Formation, it is possible that it represents a network of numerous smaller shallow waterbodies that collectively offered a diversity of environments for colonization, making it a unique deposit for examination of early freshwater diatoms.