A microfluidic chemostat for experiments with bacterial and yeast cells

Nat Methods. 2005 Sep;2(9):685-9. doi: 10.1038/nmeth784.

Abstract

Bacteria and yeast frequently exist as populations capable of reaching extremely high cell densities. With conventional culturing techniques, however, cell proliferation and ultimate density are limited by depletion of nutrients and accumulation of metabolites in the medium. Here we describe design and operation of microfabricated elastomer chips, in which chemostatic conditions are maintained for bacterial and yeast colonies growing in an array of shallow microscopic chambers. Walls of the chambers are impassable for the cells, but allow diffusion of chemicals. Thus, the chemical contents of the chambers are maintained virtually identical to those of the nearby channels with continuous flowthrough of a dynamically defined medium. We demonstrate growth of cell cultures to densely packed ensembles that proceeds exponentially in a temperature-dependent fashion, and we use the devices to monitor colony growth from a single cell and to analyze the cell response to an exogenously added autoinducer.

Publication types

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

MeSH terms

  • Bioreactors / microbiology*
  • Cell Separation / instrumentation*
  • Cell Separation / methods
  • Equipment Design
  • Equipment Failure Analysis
  • Escherichia coli / cytology*
  • Flow Cytometry / instrumentation*
  • Flow Cytometry / methods
  • Flow Injection Analysis / instrumentation*
  • Flow Injection Analysis / methods
  • Image Cytometry / instrumentation
  • Image Cytometry / methods
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods
  • Saccharomyces cerevisiae / cytology*