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Slipstreaming Mother Machine: A Microfluidic Device for Single-Cell Dynamic Imaging of Yeast

David C. Duran,Cesar A. Hernandez,Elizabeth Suesca,Ruben Acevedo, Ivon M. Acosta, Diana A. Forero, Francisco E. Rozo,Juan M. Pedraza

MICROMACHINES(2021)

Cited 4|Views20
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Abstract
The yeast Saccharomyces cerevisiae is one of the most basic model organisms for studies of aging and other phenomena such as division strategies. These organisms have been typically studied with the use of microfluidic devices to keep cells trapped while under a flow of fresh media. However, all of the existing devices trap cells mechanically, subjecting them to pressures that may affect cell physiology. There is evidence mechanical pressure affects growth rate and the movement of intracellular components, so it is quite possible that it affects other physiological aspects such as aging. To allow studies with the lowest influence of mechanical pressure, we designed and fabricated a device that takes advantage of the slipstreaming effect. In slipstreaming, moving fluids that encounter a barrier flow around it forming a pressure gradient behind it. We trap mother cells in this region and force daughter cells to be in the negative pressure gradient region so that they are taken away by the flow. Additionally, this device can be fabricated using low resolution lithography techniques, which makes it less expensive than devices that require photolithography masks with resolution under 5 mu m. With this device, it is possible to measure some of the most interesting aspects of yeast dynamics such as growth rates and Replicative Life Span. This device should allow future studies to eliminate pressure bias as well as extending the range of labs that can do these types of measurements.
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Key words
microfluidics,replicative aging,mother machine,Saccharomyces cerevisiae
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