Dielectrophoresis (DEP) is the motion of polarizable particles in the presence of nonuniform electric fields. This novel electrokinetic technique has successfully been employed in many miniaturized systems for the manipulation and detection of microbes. This review article depicts the application of dielectrophoresis for the monitoring of microorganisms in microfluidic devices for environmental applications. The research studies described here are mainly conceived for water- and air-monitoring assessments, and are classified considering the target aimed to detect, concentrate, and/or separate, including chemical and toxicant agents, and microorganisms ranging from virus to protozoa. Dielectrophoresis has also played an important role in biofilm formation studies. This review article comprises mainly studies published from 2000 to present. Even in this relatively short time frame, there have been many significant contributions of this powerful and nascent technique related to environmental monitoring; thus, unveiling its great potential for future research directions.