Organohalide respiring bacteria are difficult to enrich and isolate, which can limit research on these important organisms. The goal of this research was to develop a method to rapidly (minutes to days) enrich these organisms from a mixed culture or sediment sample. The method presented herein was based on the hypothesis that organohalide respiring bacteria would be more hydrophobic than other bacteria as a result of their niche dehalogenating hydrophobic compounds. To test this hypothesis, a method was developed to separate putative organohalide respiring bacteria at the interface between a hydrophobic organic solvent and aqueous media. This novel separation technique was tested with a sediment-free culture, a tetrachloroethene-enriched digester culture, and sediment from an uncontaminated lake.