The Eurasian freshwater tubenose goby Proterorhinus semilunaris (formerly Proterorhinus marmoratus) invaded the Laurentian Great Lakes in the 1990s, presumably via ballast water from transoceanic cargo ships. Tubenose gobies spread throughout Lake St. Clair, its tributaries, and the Detroit River system, and also are present in the Duluth-Superior harbor of Lake Superior. Using seines and bottom trawls, we collected 113 tubenose gobies between July 2007 and August 2009 at several locations in western Lake Erie. The number and range of sizes of specimens collected suggest that that tubenose gobies have become established and self-sustaining in the western basin of Lake Erie. Tubenose gobies reached maximum densities in sheltered areas with abundant macrophyte growth, which also is their common habitat in native northern Black Sea populations. The diet of tubenose gobies was almost exclusively invertebrates, suggesting dietary overlap with other benthic fishes, such as darters (Etheostoma spp. and Percina sp.), madtoms (Noturus spp.), and sculpins (Cottus spp.). A single mitochondrial DNA haplotype was identified, which is the most common haplotype found in the original colonization area in the Lake St. Clair region, suggesting a founder effect. Tubenose gobies, like round gobies Neogobius melanostomus, have early life stages that drift owing to vertical migration, which probably allowed them to spread from areas of colonization. The Lake St. Clair-Lake Erie corridor appears to have served as an avenue for them to spread to the western basin of Lake Erie, and abundance of shallow macrophyte-rich habitats may be a key factor facilitating their further expansion within Lake Erie and the remainder of the Laurentian Great Lakes.