The invention provides a droplet actuator comprising a base substrate and a top substrate separated to form a gap, wherein: (a) the base substrate comprises electrodes configured for conducting droplet operations in the gap; and (b) the top substrate comprises a first portion coupled to second portion, where the second portion comprises one or more openings establishing a fluid path extending from an exterior of the droplet actuator and into the gap. The invention also provides related methods of manufacturing the droplet actuator, methods of using the droplet actuator, and methods of loading the droplet actuator.
