Assortative mating is a reproductive strategy used by a diversity of animals, in which individuals choose a mate that shares similar characteristics. This mating strategy has the potential to promote the evolution of various sexual signals and has been a proposed mechanism driving and maintaining color variation in the anuran family Dendrobatidae. Most studies have examined this reproductive strategy in the polytypic poison frog, Oophaga pumilio, in the Bocas del Toro archipelago in Panama. Little attention, however, has been given to ancestral populations across this species’ mainland range, where dramatic color polytypism appears to lack. Additionally, most studies are exclusively experimental and investigate mate choice between allopatric populations, neglecting the behaviors of naturally occurring mates. This study observed natural mating pairs within a population of O. pumilio on mainland Costa Rica and tested the prediction that color phenotype of mating females and males would be correlated. Naturally occurring pairs were found to share similar coloration, suggesting that color assortative mating operates in nature, and in a mainland population. Our results indicate that coloration is an important trait in driving the natural mate choices of female O. pumilio, which provides valuable insight into realistic mate selection tactics of this dendrobatid frog.