In Shear’s 1986 cladistic analysis of the Ischyropsalidoidea, he described the new genus Acuclavella including four new species from the Pacific Northwest states of Washington and Idaho. Several of these species descriptions were based on very limited sample sizes. Our recent field work has increased by more than an order of magnitude both the number of specimens and known localities for Acuclavella. We use this new material to interpret species limits in Acuclavella using morphometric analyses and DNA sequence data from four gene regions. We sequence for the first time the protein-coding homolog of the WNT2 gene for phylogenetic reconstruction in Opiliones. Our multi-locus phylogeny corroborates a sister relationship between Acuclavella and Ceratolasma, as hypothesized using morphology by Shear (1986). Within Acuclavella, morphometric clusters and reciprocal allelic monophyly allows recognition of three additional species: Acuclavella leonardi sp. n., A. sheari sp. n., and A. makah sp. n. This work also describes the previously unknown male of Acuclavella quattuor, from specimens collected at the type locality. Our research identifies a number of novel morphologies for Acuclavella, including females with four pairs of spines, individuals with three pairs of spines on scute areas I-III, and a population with two pairs of spines disjunct from A. quattuor, which was diagnosed with this spination character. We were unable to assign these populations to existing species, and conservatively do not yet recognize them as new. Intrageneric morphometrics and phylogenetic inference in Acuclavella were often concordant. However, we demonstrate that species delimitation signal would not be detected if only a single line of evidence were utilized.