Long-term imaging of Caenorhabditis elegans using nanoparticle-mediated immobilization

PLoS One. 2013;8(1):e53419. doi: 10.1371/journal.pone.0053419. Epub 2013 Jan 3.

Abstract

One advantage of the nematode Caenorhabditis elegans as a model organism is its suitability for in vivo optical microscopy. Imaging C. elegans often requires animals to be immobilized to avoid movement-related artifacts. Immobilization has been performed by application of anesthetics or by introducing physical constraints using glue or specialized microfluidic devices. Here we present a method for immobilizing C. elegans using polystyrene nanoparticles and agarose pads. Our technique is technically simple, does not expose the worm to toxic substances, and allows recovery of animals. We evaluate the method and show that the polystyrene beads increase friction between the worm and agarose pad. We use our method to quantify calcium transients and long-term regrowth in single neurons following axotomy by a femtosecond laser.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axotomy / instrumentation
  • Axotomy / methods
  • Caenorhabditis elegans / metabolism*
  • Calcium Signaling
  • Immobilization / instrumentation
  • Immobilization / methods*
  • Microfluidic Analytical Techniques
  • Microscopy / methods*
  • Movement
  • Nanoparticles / chemistry*
  • Nanotechnology / methods*
  • Neurons / metabolism
  • Polystyrenes / chemistry
  • Sepharose / chemistry

Substances

  • Polystyrenes
  • Sepharose

Grant support

EK was partly supported by the Penn Undergraduate Research Mentorship program. CVG was partly supported by the Massachusetts Life Science Center New Investigator Matching Grant. (http://www.masslifesciences.com/grants/invest.html) No additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.