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High-Resolution C. elegans Imaging Across All Larval Stages.

Simon Berger1, Silvan Spiri2, Andrew deMello3

  • 1Department of Molecular Life Sciences, University Zürich; simon.berger@mls.uzh.ch.

Journal of Visualized Experiments : Jove
|June 9, 2025
PubMed
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A new microfluidic method immobilizes Caenorhabditis elegans (C. elegans) for live imaging without harming development. This technique preserves worm orientation and identity during time-lapse microscopy, improving biological studies.

Area of Science:

  • * Developmental Biology
  • * Microscopy Techniques
  • * Model Organisms

Background:

  • * Caenorhabditis elegans (C. elegans) is a key model organism due to its invariant cell lineage, transparency, and genetic tractability.
  • * In vivo microscopy in C. elegans requires immobilization, but traditional methods can negatively impact animal development, especially in time-lapse studies.
  • * Existing microfluidic immobilization strategies have limitations that hinder simultaneous imaging across larval stages and long-term observation.

Purpose of the Study:

  • * To introduce a novel microfluidic imaging method for immobilizing C. elegans.
  • * To overcome the limitations of traditional agar-pad and other microfluidic immobilization techniques.
  • * To enable high-resolution, time-lapse live imaging of C. elegans while preserving worm viability and developmental timing.

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Main Methods:

  • * Development of a microfluidic trap channel array with geometry optimized for stable worm orientation and accommodation of growth/molting.
  • * Implementation of an active hydraulic valve system for precise, temporary immobilization against the cover glass during image acquisition.
  • * Protocol detailing setup and operation for simultaneous live imaging of various C. elegans larval stages.

Main Results:

  • * The novel microfluidic method successfully immobilizes C. elegans larvae and adults for high-resolution imaging.
  • * Worm orientation and identity are preserved over time, facilitating longitudinal studies.
  • * Minimal adverse effects on C. elegans viability and developmental timing were observed compared to traditional methods.

Conclusions:

  • * The developed microfluidic imaging approach offers a significant improvement for live C. elegans studies.
  • * This method enables advanced microscopy applications, including long-term time-lapse imaging, without compromising animal welfare.
  • * The technique supports diverse microscopy-based research in C. elegans, enhancing its utility as a model organism.