Femtosecond laser axotomy in Caenorhabditis elegans and collateral damage assessment using a combination of linear and nonlinear imaging techniques

PLoS One. 2013;8(3):e58600. doi: 10.1371/journal.pone.0058600. Epub 2013 Mar 6.

Abstract

In this work highly localized femtosecond laser ablation is used to dissect single axons within a living Caenorhabditis elegans (C. elegans). We present a multimodal imaging methodology for the assessment of the collateral damage induced by the laser. This relies on the observation of the tissues surrounding the targeted region using a combination of different high resolution microscopy modalities. We present the use of Second Harmonic Generation (SHG) and Polarization Sensitive SHG (PSHG) to determine damage in the neighbor muscle cells. All the above is done using a single instrument: multimodal microscopy setup that allows simultaneous imaging in the linear and non-linear regimes and femtosecond-laser ablation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axotomy / adverse effects
  • Axotomy / methods*
  • Caenorhabditis elegans
  • Laser Therapy / adverse effects
  • Laser Therapy / methods*
  • Microscopy, Electron, Transmission
  • Microscopy, Polarization / methods*

Grant support

This work is supported by the Generalitat de Catalunya, the Spanish government grants TEC2009-09698 and the FAST DOT EU project. Authors also acknowledge the Laser Lab Europe (optobio) and the Photonics4Life network. This research has been partially supported by Fundació Cellex Barcelona and has been conducted at ICFO's “Super-Resolution Light Microscopy and Nanoscopy Facility” (SLN@ICFO). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.