Native freshwater mussels (Unionidae) are a diverse and imperiled group of organisms with significant conservation value. The Tonawanda Creek basin has one of the most diverse assemblages of unionids in western New York. Previous surveys have been conducted in Tonawanda Creek and its tributaries to characterize this ecologically significant assemblage. However, most of these surveys were qualitative and do not provide density estimates. They are also less likely to detect juvenile mussels (< 20 mm in length), which may be more sensitive to certain stressors. To better understand the recruitment statuses and habitat preferences of mussel populations in western New York, we conducted quantitative mussel surveys at stream locations with known mussel presence. Additionally, we assessed habitat quality, including substrate type and water chemistry, to better understand the conditions required for successful mussel recruitment. We surveyed six sites across three creeks (Tonawanda, Ellicott, and Ransom Creeks) and detected 16 live mussel species and an additional 3 species as spent shells. While Tonawanda Creek itself accounted for the majority of this diversity, three of the live species were exclusively found in Ellicott Creek. Mussel densities ranged from 0.4 to 16.27 mussels/m2, and sites with higher percent fine sediment (> 50 %) and higher water hardness (> 480 CaCO3) had the lowest mussel densities. We detected recent recruitment at all but one of six sites. Our surveys show that most of the largest mussel beds in Tonawanda Creek and its tributaries have retained the majority of their diversity and are actively recruiting. However, declines in catch per unit effort and species richness could indicate that these populations remain vulnerable. By assessing the habitat preferences and recruitment statuses within a speciose mussel assemblage, these surveys will facilitate the conservation of this highly imperiled group of animals.