Single-Cell Cultivation in Microfluidic Devices
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.
KeywordsImaging Live cell Microfluidic Single cell
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