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A microfluidic array for large-scale ordering and orientation of embryos

Nature Methods volume 8, pages 171–176 (2011)Cite this article

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Abstract

Quantitative studies of embryogenesis require the ability to monitor pattern formation and morphogenesis in large numbers of embryos, at multiple time points and in diverse genetic backgrounds. We describe a simple approach that greatly facilitates these tasks for Drosophila melanogaster embryos, one of the most advanced models of developmental genetics. Based on passive hydrodynamics, we developed a microfluidic embryo-trap array that can be used to rapidly order and vertically orient hundreds of embryos. We describe the physical principles of the design and used this platform to quantitatively analyze multiple morphogen gradients in the dorsoventral patterning system. Our approach can also be used for live imaging and, with slight modifications, could be adapted for studies of pattern formation and morphogenesis in other model organisms.

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Figure 1: Microfluidic embryo-trap array for high-throughput arraying of vertically oriented Drosophila embryos.
Figure 2: Operating principles of the embryo-trap array.
Figure 3: Spatial extent of the Dl gradient.
Figure 4: Quantitative characterization of signal transduction and morphogen gradients in dorsoventral patterning.
Figure 5: Live imaging of embryos using the embryo array.

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Acknowledgements

We acknowledge A. Boettiger and M. Levine (University of California, Berkeley) for the antibody to Twist, M. Zhan for technical assistance, A. Boettiger, A. Erives, M. Levine, J. Lippincott-Schwartz, C. Rushlow, M. Serpe and R. Steward for helpful discussions, and M. Osterfield for assistance with live imaging. This work was supported by National Science Foundation (DBI-0649833 to H.L.) and National Institutes of Health grants NS058465 (to H.L.) and GM078079 (to S.Y.S.). H.L. is supported by a Sloan Foundation Research Fellowship and a DuPont Young Professor grant.

Author information

Author notes

  1. Kwanghun Chung and Yoosik Kim: These authors contributed equally to this work.

Authors and Affiliations

  1. School of Chemical and Biomolecular Engineering, and Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia, USA

    Kwanghun Chung, Emily Gong & Hang Lu

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

    Yoosik Kim, Jitendra S Kanodia & Stanislav Y Shvartsman

Authors
  1. Kwanghun Chung
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  2. Yoosik Kim
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  3. Jitendra S Kanodia
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  4. Emily Gong
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  5. Stanislav Y Shvartsman
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  6. Hang Lu
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Contributions

K.C., E.G. and H.L. designed, fabricated and tested the device. Y.K. tested the device and performed imaging. J.S.K. wrote the image processing and statistical analysis programs for gradient quantification. K.C., Y.K., S.Y.S. and H.L. designed the experiments and wrote the paper.

Corresponding author

Correspondence to Hang Lu.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1– 4 (PDF 504 kb)

Supplementary Video 1

Drosophila embryo trapping. This movie shows trapping of the embryos from an embryo suspension. (MOV 2964 kb)

Supplementary Video 2

Contraction of the traps. This movie shows automatic contraction of the traps resulting from the loading pressure being decreased from 6 psi to 0 psi. Notice that embryos in the traps are not secured. (MOV 336 kb)

Supplementary Video 3

Live imaging: early embryo. This movie shows consecutive nuclear divisions in the early embryo. (MOV 3405 kb)

Supplementary Video 4

Live imaging: ventral invagination. This movie shows consecutive nuclear divisions in an embryo undergoing ventral invagination. (MOV 994 kb)

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Chung, K., Kim, Y., Kanodia, J. et al. A microfluidic array for large-scale ordering and orientation of embryos. Nat Methods 8, 171–176 (2011). https://doi.org/10.1038/nmeth.1548

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