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Here we present a versatile system for milliliter-scale perfusion culture of adherent cells that can be built using basic tools, based on a readily available one-well culture plate (84 cm 2 culture area). Media composition and flow paths can be programmatically controlled via USB serial interface using the FETbox hardware controller and associated PlateFlo Python package. The FETbox can control up to five high current 12 V devices such as common pinch valves, solenoids, and DC motor peristaltic pumps. It was designed to be easily customized with built-in accommodation for additional electronic components (e.g. analog sensors and input), use of the ubiquitous Arduino Nano platform, and easily expanded serial communication protocol. Multiple FETboxes can be used in parallel for additional devices. Applications of the PlateFlo system include perfusion culture of laboratory experiments requiring large cell numbers including genome-scale genetic screens and proteomics, as well as novel perfusion schemes including dynamic media conditions and sequential cell culture.

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