Movie S1.

SNDA loading. The SNDA–AST device is loaded using a single step injection of a two-plug solution using a conventional 10 µL laboratory pipette. The first, or bottommost, plug is a ∼1.6-µL bacterial suspension of 5×105 cfu/mL supplemented with 10% resazurin. The second, or topmost, plug contains ∼3 µL of FC-40 oil. The two-plug solution is achieved simply by aspirating the respective liquids sequentially. When injected, the bacterial suspension in the first plug fills the wells of the SNDA–AST device in a passive manner because the air is able to escape easily through gap restrictions in the device. Next, the introduction of the second oil phase into the main channel effectively seals the chambers by separating well volumes from one another via removing connecting fluid from the main channel. The fluorinated structure of the oil allows for high oxygen solubility and aids in delivering oxygen to the bacterial cell culture in the wells.