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The CellClamper: A Convenient Microfluidic Device for Time-Lapse Imaging of Yeast

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Genome Instability

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1672))

Abstract

Time-lapse fluorescence imaging of yeast cells allows the study of multiple fluorescent targets in single cells, but is often hampered by the tedious cultivation using agar pads or glass bottom wells. Here, we describe the fabrication and operation of a microfluidic device for long-term imaging of yeast cells under constant or changing media conditions. The device allows acquisition of high quality images as cells are fixed in a two-dimensional imaging plane. Four yeast strains can be analyzed simultaneously over several days while up to four different media can be flushed through the chip. The microfluidic device does not rely on specialized equipment for its operation. To illustrate the use of the chip in DNA damage research, we show how common readouts for DNA damage or genomic instability behave upon induction with genotoxic chemicals (MMS, HU) or induction of a single double-strand break using induced CRISPR-Cas9 expression.

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Acknowledgements

This work was supported by the FP7 grant no 28995 ITN ISOLATE and the Ambizione grant 142440 of the Swiss National Science Foundation to Olivier Frey. We thank Andreas Hierlemann and Jörg Stelling for their support. We acknowledge Mjriam Bächler and Andreas Cuny for testing the microfluidic device and Jennifer Ewald and Tania Roberts for critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland

    Gregor W. Schmidt, Olivier Frey & Fabian Rudolf

Authors
  1. Gregor W. Schmidt
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  2. Olivier Frey
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  3. Fabian Rudolf
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Corresponding author

Correspondence to Fabian Rudolf .

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Editors and Affiliations

  1. Dipartimento di Bioscienze, Università degli Studi di Milano, Milano, Milano, Italy

    Marco Muzi-Falconi

  2. Donnelly Centre and Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada

    Grant W Brown

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Schmidt, G.W., Frey, O., Rudolf, F. (2018). The CellClamper: A Convenient Microfluidic Device for Time-Lapse Imaging of Yeast. In: Muzi-Falconi, M., Brown, G. (eds) Genome Instability. Methods in Molecular Biology, vol 1672. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7306-4_36

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  • DOI: https://doi.org/10.1007/978-1-4939-7306-4_36

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7305-7

  • Online ISBN: 978-1-4939-7306-4

  • eBook Packages: Springer Protocols

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