The ability of individuals within a population to survive and thrive is highly dependent upon the maintenance of genetic variation and phenotypic diversity, thereby ensuring adaptation to dynamic environments. A fundamental method of maintaining such variation is through a negative assortative mating strategy, in which individuals would be expected to reproductively select members of the opposite sex that exhibit dissimilar phenotypes. Employing three uniform body color morphs, red, yellow and blue, of the platyfish (Xiphophorus maculatus), this study was designed to investigate whether X. maculatus females would preferentially be attracted to males exhibiting an alternative color, thereby enabling an examination of the effect of male body coloration on mate choice by adult females. Mate choice was determined based on the initial preference of each female, as well as the amount of time females spent associating with each male. Initial preferences were analyzed using a binomial distribution test, and overall preference data using Wilcoxon signed rank tests. Red females initially selected for dissimilar colored males, and spent a significantly larger amount of time associating with blue and yellow males, as did yellow females with red and blue males. Blue females initially selected and spent a significantly larger amount of time associating with red males but, interestingly, showed no selective preference between blue and yellow males. In these experimental trials, the overall strong mate selection exhibited by female platyfish for males of dissimilar coloration is suggestive of a negative assortative mating strategy and provides evidence for the maintenance of color polymorphism in nature populations.