title: Design and use of multiplexed chemostat arrays.
abstract: Chemostats are continuous culture systems in which cells are grown in a tightly controlled, chemically constant environment where culture density is constrained by limiting specific nutrients.(1,2) Data from chemostats are highly reproducible for the measurement of quantitative phenotypes as they provide a constant growth rate and environment at steady state. For these reasons, chemostats have become useful tools for fine-scale characterization of physiology through analysis of gene expression(3-6) and other characteristics of cultures at steady-state equilibrium.(7) Long-term experiments in chemostats can highlight specific trajectories that microbial populations adopt during adaptive evolution in a controlled environment. In fact, chemostats have been used for experimental evolution since their invention.(8) A common result in evolution experiments is for each biological replicate to acquire a unique repertoire of mutations.(9-13) This diversity suggests that there is much left to be discovered by performing evolution experiments with far greater throughput. We present here the design and operation of a relatively simple, low cost array of miniature chemostats-or ministats-and validate their use in determination of physiology and in evolution experiments with yeast. This approach entails growth of tens of chemostats run off a single multiplexed peristaltic pump. The cultures are maintained at a 20 ml working volume, which is practical for a variety of applications. It is our hope that increasing throughput, decreasing expense, and providing detailed building and operation instructions may also motivate research and industrial application of this design as a general platform for functionally characterizing large numbers of strains, species, and growth parameters, as well as genetic or drug libraries.
authors: Department of Genome Sciences, University of Washington, WA, USA.
authors: Aaron W
authors: AW
authors: Miller
authors: Corrie
authors: C
authors: Befort
authors: Emily O
authors: EO
authors: Kerr
authors: Maitreya J
authors: MJ
authors: Dunham
mesh_terms: D002468:Cell Physiological Phenomena; D003584:Cytological Techniques / Q000191:economics / Q000295:instrumentation* / Q000379:methods*; D004718:Saccharomycetales / Q000166:cytology