Devices and methods for fluid actuation

A digital microfluidic and analyte detection device, comprising:

• a first substrate and a second substrate aligned generally parallel to each other with a gap defined therebetween in side view, at least one of the first substrate and the second substrate including:
• a first electrode array,
• a second electrode array spaced from and in electrical communication with the first electrode array, and
• an interstitial area defined between the first electrode array and the second electrode array;
• an analyte detection device defined in at least one of the first substrate and the second substrate, wherein at least one of the first electrode array and the second electrode array is configured to generate electrical actuation forces within an actuation area to urge at least one droplet within the gap along the at least one of the first substrate and the second substrate to the analyte detection device;
• and
• at least one spacer disposed in the interstitial area to maintain the gap between the first substrate and the second substrate.