Building a morbidostat: an automated continuous-culture device for studying bacterial drug resistance under dynamically sustained drug inhibition

Nat Protoc. 2013 Mar;8(3):555-67. doi: 10.1038/nprot.nprot.2013.021. Epub 2013 Feb 21.

Abstract

We present a protocol for building and operating an automated fluidic system for continuous culture that we call the 'morbidostat'. The morbidostat is used to follow the evolution of microbial drug resistance in real time. Instead of exposing bacteria to predetermined drug environments, the morbidostat constantly measures the growth rates of evolving microbial populations and dynamically adjusts drug concentrations inside culture vials in order to maintain a constant drug-induced inhibition. The growth rate measurements are done using an optical detection system that is based on measuring the intensity of back-scattered light from bacterial cells suspended in the liquid culture. The morbidostat can additionally be used as a chemostat or a turbidostat. The whole system can be built from readily available components within 2-3 weeks by biologists with some electronics experience or engineers familiar with basic microbiology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Bacillus subtilis / drug effects
  • Bacillus subtilis / growth & development
  • Bacteria / drug effects*
  • Bacteria / genetics
  • Bacteria / growth & development
  • Cell Culture Techniques / instrumentation*
  • Drug Resistance, Bacterial*
  • Escherichia coli / drug effects
  • Escherichia coli / growth & development
  • Evolution, Molecular
  • Microbial Sensitivity Tests / instrumentation
  • Microbial Sensitivity Tests / methods

Substances

  • Anti-Bacterial Agents