Single-Cell Cultivation in Microfluidic Devices

Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Live-cell imaging is a powerful technique for investigating physiological aspects of growing cells. Monitoring a large number of cells is desired to collect data of statistical significance, from which one can address key biological questions. However, traditional cell-immobilization techniques (i.e., agar pads) limit the possibility of large data collection as colonies can rapidly outgrow a field of view and use up available nutrients. Here, we present a protocol for fabrication of a microfluidic device, the “mother machine”, that allows long-term growth experiment on rod-shaped cells. In a mother machine, continuous flow of growth medium provides nutrients for growing cells and clears the device of excess cells. The device is assembled on a glass-bottom dish from which we can perform multi-point time-lapse microscopy. The design has approximately 1,000 growth channels per device, which allow us capturing data on more than 105 individual cells in one single experiment.

Keywords

Imaging Live cell Microfluidic Single cell 

References

  1. 1.
    Wang P, Robert L, Pelletier J, Dang WL, Taddei F, Wright A, Jun S (2010) Robust growth of Escherichia coli. Curr Biol 20(12):1099–1103CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Johnston ID, McCluskey DK, Tan CKL, Tracey MC (2014) Mechanical characterization of bulk Sylgard 184 for microfluidics and microengineering. J Micromech Microeng 24(3):035017-1–035017-7Google Scholar
  3. 3.
    Ito T, Okazaki S (2000) Pushing the limits of lithography. Nature 406(6799):1027–1031CrossRefPubMedGoogle Scholar
  4. 4.
    Lee JN, Park C, Whitesides GM (2003) Solvent compatibility of poly(dimethylsiloxane)-based microfluidic devices. Anal Chem 75(23):6544–6554CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.Department of PhysicsUniversity of California San DiegoLa JollaUSA
  2. 2.Section of Molecular Biology, Division of Biological ScienceUniversity of California San DiegoLa JollaUSA

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