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Microfluidic trap array for massively parallel imaging of Drosophila embryos

Nature Protocols volume 8, pages 721–736 (2013)Cite this article

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Abstract

Here we describe a protocol for the fabrication and use of a microfluidic device to rapidly orient >700 Drosophila embryos in parallel for end-on imaging. The protocol describes master microfabrication (∼1 d), polydimethylsiloxane molding (few hours), system setup and device operation (few minutes) and imaging (depending on application). Our microfluidics-based approach described here is one of the first to facilitate rapid orientation for end-on imaging, and it is a major breakthrough for quantitative studies on Drosophila embryogenesis. The operating principle of the embryo trap is based on passive hydrodynamics, and it does not require direct manipulation of embryos by the user; biologists following the protocol should be able to repeat these procedures. The compact design and fabrication materials used allow the device to be used with traditional microscopy setups and do not require specialized fixtures. Furthermore, with slight modification, this array can be applied to the handling of other model organisms and oblong objects.

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Figure 1: Device and protocol overview.
Figure 2: Equipment setup.
Figure 3: Microfabrication of SU-8 master.
Figure 4: Common problems.
Figure 5: Fabrication of the PDMS device.
Figure 6: Device loading.

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Acknowledgements

This work was supported by the US National Science Foundation (DBI-0649833 to H.L., EFRI 1136913 to S.Y.S. and H.L.) and the US National Institutes of Health (grant nos. NS058465 to H.L. and GM078079 to S.Y.S.). H.L. is a DuPont Young Professor and a Sloan Research Fellow. We thank J. Ding for technical assistance.

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Authors and Affiliations

  1. School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA

    Thomas J Levario & Hang Lu

  2. Interdisciplinary Program in Bioengineering, Institute of Biosciences and Bioengineering, Georgia Institute of Technology, Atlanta, Georgia, USA

    Mei Zhan & Hang Lu

  3. Department of Chemical and Biological Engineering and Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA

    Bomyi Lim & Stanislav Y Shvartsman

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  1. Thomas J Levario
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Contributions

T.J.L. and M.Z. fabricated the devices and tested the protocol. H.L. and S.Y.S. oversaw the project. T.J.L. wrote the paper. All authors (T.J.L., M.Z.,B.L., S.Y.S. and H.L.) contributed to the design of the experiments and edited the manuscript.

Corresponding author

Correspondence to Hang Lu.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Figure 1

Microfluidic device layout and specifications (PDF 465 kb)

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Levario, T., Zhan, M., Lim, B. et al. Microfluidic trap array for massively parallel imaging of Drosophila embryos. Nat Protoc 8, 721–736 (2013). https://doi.org/10.1038/nprot.2013.034

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