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This dataset contains species richness data for epiphytic diatom communities collected from weir ponds in seven headwater streams within the Hubbard Brook Experimental Forest (HBEF) in New Hampshire between 2018 and 2021. Diatom samples were gathered using artificial bryophyte substrates, deployed in weir ponds to mimic natural diatom habitats. Species richness was quantified by identifying diatom taxa to the lowest possible taxonomic level, with 86 taxa spanning 43 genera recorded. This dataset represents the first comprehensive classification of diatom communities at HBEF, providing a baseline for future studies in this ecosystem. Environmental variables, including light availability, dissolved organic carbon, total dissolved nitrogen, and pH, were concurrently measured to assess their influence on diatom community composition. The light (lux) data used in this study is openly available in the EDI Data Portal at https://doi.org/10.6073/pasta/0f40b75b299494d736645d940fa2b5a4. The chlorophyll-a data and analysis methodology are available at https://doi.org/10.6073/pasta/7fa32d94240fc7780d62cb7e65eafdb2. Reach characteristics were sourced from the EDI Data Portal at https://doi.org/10.6073/pasta/3e4b95149245341d522383bba51de7c7. This study provides valuable insights into the relationships between environmental factors and diatom diversity in northern hardwood forest streams, aiding ecological monitoring and bioindicator studies. These data were gathered as part of the Hubbard Brook Ecosystem Study (HBES). The HBES is a collaborative effort at the Hubbard Brook Experimental Forest, which is operated and maintained by the USDA Forest Service, Northern Research Station.