Protocol

Hydrocarbon and Lipid Microbiology Protocols

Part of the series Springer Protocols Handbooks pp 5-16

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Single-Cell Cultivation in Microfluidic Devices

  • Sattar Taheri-AraghiAffiliated withDepartment of Physics, University of California San Diego
  • , Suckjoon JunAffiliated withDepartment of Physics, University of California San DiegoSection of Molecular Biology, Division of Biological Science, University of California San Diego Email author 

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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