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Femtosecond laser nanoaxotomy lab-on-a-chip for in vivo nerve regeneration studies

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Abstract

A thorough understanding of nerve regeneration in Caenorhabditis elegans requires performing femtosecond laser nanoaxotomy while minimally affecting the worm. We present a microfluidic device that fulfills such criteria and can easily be automated to enable high-throughput genetic and pharmacological screenings. Using the 'nanoaxotomy' chip, we discovered that axonal regeneration occurs much faster than previously described, and notably, the distal fragment of the severed axon regrows in the absence of anesthetics.

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Figure 1: The nanoaxotomy lab-on-a-chip.
Figure 2: Time-lapse imaging of axonal recovery on a chip.

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Acknowledgements

We thank C. Bargmann and M. Goodman for valuable discussions. This work was supported by grants from the US National Institutes of Health (NS058646 and NS060129) and the National Science Foundation (BES-0548673).

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Authors and Affiliations

  1. Department of Mechanical Engineering, The University of Texas at Austin, 204 E. Dean Keeton Street, Austin, 78712, Texas, USA

    Samuel X Guo, Frederic Bourgeois, Nicholas J Durr & Adela Ben-Yakar

  2. Department of Mechanical Engineering, University of Michigan, 2350 Hayward Street, Ann Arbor, 48109, Michigan, USA

    Trushal Chokshi & Nikos Chronis

  3. Queensland Brain Institute, University of Queensland, Brisbane, QLD 4072, Australia

    Massimo A Hilliard

Authors
  1. Samuel X Guo
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  2. Frederic Bourgeois
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  3. Trushal Chokshi
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  4. Nicholas J Durr
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  5. Massimo A Hilliard
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  6. Nikos Chronis
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  7. Adela Ben-Yakar
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Contributions

N.C. and A.B. designed the device; T.C. fabricated the microfluidic chip; S.X.G. and N.D. performed two-photon microscopy; F.B., M.H. and A.B. analyzed data; S.X.G., F.B. and A.B. designed experiments; S.X.G. and F.B. performed experiments; F.B. and A.B. wrote the paper.

Corresponding author

Correspondence to Adela Ben-Yakar.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3, Supplementary Tables 1–2, Supplementary Methods (PDF 957 kb)

Supplementary Video 1

Immobilization of a worm in the microfluidic trap. (MOV 262 kb)

Supplementary Video 2

Laser nanoaxotomy of a worm on a chip. (MOV 2873 kb)

Supplementary Video 3

Behavioral assay of a worm following nanoaxotomy on the branches of all of its PLM and ALM processes. (MOV 350 kb)

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Guo, S., Bourgeois, F., Chokshi, T. et al. Femtosecond laser nanoaxotomy lab-on-a-chip for in vivo nerve regeneration studies. Nat Methods 5, 531–533 (2008). https://doi.org/10.1038/nmeth.1203

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  • DOI: https://doi.org/10.1038/nmeth.1203

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