Abstract
Recent advances in gene editing are enabling the engineering of cells with an unprecedented level of scale. To capitalize on this opportunity, new methods are needed to accelerate the different steps required to manufacture and handle engineered cells. Here, we describe the development of an integrated software and hardware platform to automate Fluorescence-Activated Cell Sorting (FACS), a central step for the selection of cells displaying desired molecular attributes. Sorting large numbers of samples is laborious, and, to date, no automated system exists to sequentially manage FACS samples, likely owing to the need to tailor sorting conditions (“gating”) to each individual sample. Our platform is built around a commercial instrument and integrates the handling and transfer of samples to and from the instrument, autonomous control of the instrument’s software, and the algorithmic generation of sorting gates, resulting in walkaway functionality. Automation eliminates operator errors, standardizes gating conditions by eliminating operator-to-operator variations, and reduces hands-on labor by 93%. Moreover, our strategy for automating the operation of a commercial instrument control software in the absence of an Application Program Interface (API) exemplifies a universal solution for other instruments that lack an API. Our software and hardware designs are fully open-source and include step-by-step build documentation to contribute to a growing open ecosystem of tools for high-throughput cell biology.
Competing Interest Statement
The authors of this manuscript have the following competing interests: The SH800S cell sorter that is part of the automation system is a commercial product sold by Sony Corporation. Fumitaka Otsuka and Yoshitsugu Sakai are employees of Sony Corporation. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. This does not alter our ability to share data and materials.
Footnotes
Updated statement about competing interests. In the Introduction, clarified the state of commercial instrumentation. Clarified issues around testing with different cell lines, with the comparison with manual sorting, and with throughput, all in the Results section. Updated the discussion section to better address manual vs automated gate generation questions (replaced S3 Fig with S4 File). Added the cost of the automation system to the Materials and Methods section.
