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Construction and use of a microfluidic dissection platform for long-term imaging of cellular processes in budding yeast

Nature Protocols volume 8, pages 1019–1027 (2013)Cite this article

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Abstract

This protocol describes the production and operation of a microfluidic dissection platform for long-term, high-resolution imaging of budding yeast cells. At the core of this platform is an array of micropads that trap yeast cells in a single focal plane. Newly formed daughter cells are subsequently washed away by a continuous flow of fresh culture medium. In a typical experiment, 50–100 cells can be tracked during their entire replicative lifespan. Apart from aging-related research, the microfluidic platform can also be a valuable tool for other studies requiring the monitoring of single cells over time. Here we provide step-by-step instructions on how to fabricate the silicon wafer mold, how to produce and operate the microfluidic device and how to analyze the obtained data. Production of the microfluidic dissection platform and setting up an aging experiment takes ∼7 h.

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Figure 1: Removal of daughter cells is essential for lifelong tracking of aging yeast cells.
Figure 2: Design of the silicon wafer mold.
Figure 3: Illustration of the most important steps in the protocol.
Figure 4: Fabrication of the silicon wafer mold and microfluidic dissection platform.
Figure 5: Operation of the microfluidic dissection platform.

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Change history

  • 13 May 2013

     In the version of this article initially published online, the author name for Javier González was misspelled as Gonzáles. The error has been corrected in the print, PDF and HTML versions of the article.

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Acknowledgements

We would like to thank M. Peter and U. Sauer for generously allowing us the use of the microscope; FIRST-CLA at ETH Zurich for the clean room facility during the development of this protocol; L. Lee for initial discussions; and N. Thayer, A. Papagiannakis and A. Meinema for helpful comments on the manuscript. Funding from the NWO-funded Groningen Systems Biology Center for Energy Metabolism and Ageing is acknowledged.

Author information

Author notes

  1. Daphne H E W Huberts and Sung Sik Lee: These authors contributed equally to this work.

Authors and Affiliations

  1. Molecular Systems Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands.,

    Daphne H E W Huberts & Matthias Heinemann

  2. Institute of Biochemistry, Zurich, Switzerland

    Sung Sik Lee

  3. Probability and Statistics Research Group, Johann Bernoulli Institute of Mathematics and Computer Science, University of Groningen, Groningen, The Netherlands.,

    Javier González

  4. Department for the Biology of Ageing, European Research Institute for the Biology of Ageing, University Medical Centre Groningen, University of Groningen, Groningen, The Netherlands.,

    Georges E Janssens

  5. Department of Health Sciences and Technology, Zurich, Switzerland

    Ima Avalos Vizcarra

Authors
  1. Daphne H E W Huberts
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  2. Sung Sik Lee
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  3. Javier González
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  4. Georges E Janssens
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  5. Ima Avalos Vizcarra
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  6. Matthias Heinemann
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Contributions

S.S.L. invented the microfluidic dissection method; I.A.V., G.E.J. and D.H.E.W.H. helped to further develop the protocol; J.G. introduced the statistical analyses; M.H. supervised the project; S.S.L., D.H.E.W.H. and M.H. wrote this protocol.

Corresponding author

Correspondence to Matthias Heinemann.

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The authors declare no competing financial interests.

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Huberts, D., Sik Lee, S., González, J. et al. Construction and use of a microfluidic dissection platform for long-term imaging of cellular processes in budding yeast. Nat Protoc 8, 1019–1027 (2013). https://doi.org/10.1038/nprot.2013.060

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