Designing Automated, High-throughput, Continuous Cell Growth Experiments Using eVOLVER

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dc.citation.articleNumbere59652
dc.citation.journalTitleJournal of Visualized Experiments
dc.citation.volumeNumber147
dc.contributor.authorHeins, Zachary J.
dc.contributor.authorMancuso, Christopher P.
dc.contributor.authorKiriakov, Szilvia
dc.contributor.authorWong, Brandon G.
dc.contributor.authorBashor, Caleb J.
dc.contributor.authorKhalil, Ahmad S.
dc.date.accessioned2019-11-22T16:19:56Z
dc.date.available2019-11-22T16:19:56Z
dc.date.issued2019
dc.description.abstractContinuous culture methods enable cells to be grown under quantitatively controlled environmental conditions, and are thus broadly useful for measuring fitness phenotypes and improving our understanding of how genotypes are shaped by selection. Extensive recent efforts to develop and apply niche continuous culture devices have revealed the benefits of conducting new forms of cell culture control. This includes defining custom selection pressures and increasing throughput for studies ranging from long-term experimental evolution to genome-wide library selections and synthetic gene circuit characterization. The eVOLVER platform was recently developed to meet this growing demand: a continuous culture platform with a high degree of scalability, flexibility, and automation. eVOLVER provides a single standardizing platform that can be (re)-configured and scaled with minimal effort to perform many different types of high-throughput or multi-dimensional growth selection experiments. Here, a protocol is presented to provide users of the eVOLVER framework a description for configuring the system to conduct a custom, large-scale continuous growth experiment. Specifically, the protocol guides users on how to program the system to multiplex two selection pressures - temperature and osmolarity - across many eVOLVER vials in order to quantify fitness landscapes of Saccharomyces cerevisiae mutants at fine resolution. We show how the device can be configured both programmatically, through its open-source web-based software, and physically, by arranging fluidic and hardware layouts. The process of physically setting up the device, programming the culture routine, monitoring and interacting with the experiment in real-time over the internet, sampling vials for subsequent offline analysis, and post experiment data analysis are detailed. This should serve as a starting point for researchers across diverse disciplines to apply eVOLVER in the design of their own complex and high-throughput cell growth experiments to study and manipulate biological systems.
dc.identifier.citationHeins, Zachary J., Mancuso, Christopher P., Kiriakov, Szilvia, et al.. "Designing Automated, High-throughput, Continuous Cell Growth Experiments Using eVOLVER." <i>Journal of Visualized Experiments,</i> 147, (2019) JoVE: https://doi.org/10.3791/59652.
dc.identifier.digitaljove-protocol-59652
dc.identifier.doihttps://doi.org/10.3791/59652
dc.identifier.urihttps://hdl.handle.net/1911/107717
dc.language.isoeng
dc.publisherJoVE
dc.rightsCreative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/3.0/
dc.titleDesigning Automated, High-throughput, Continuous Cell Growth Experiments Using eVOLVER
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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