Optogenetic manipulation of neural activity in freely moving Caenorhabditis elegans

Andrew M. Leifer, Christopher Fang-Yen, Marc Gershow, Mark J. Alkema, Aravinthan D.T. Samuel

Research output: Contribution to journalArticlepeer-review

244 Scopus citations

Abstract

We present an optogenetic illumination system capable of real-time light delivery with high spatial resolution to specified targets in freely moving Caenorhabditis elegans. A tracking microscope records the motion of an unrestrained worm expressing channelrhodopsin-2 or halorhodopsin in specific cell types. Image processing software analyzes the worm's position in each video frame, rapidly estimates the locations of targeted cells and instructs a digital micromirror device to illuminate targeted cells with laser light of the appropriate wavelengths to stimulate or inhibit activity. Because each cell in an unrestrained worm is a rapidly moving target, our system operates at high speed (∼50 frames per second) to provide high spatial resolution (∼30 μm). To test the accuracy, flexibility and utility of our system, we performed optogenetic analyses of the worm motor circuit, egg-laying circuit and mechanosensory circuits that have not been possible with previous methods.

Original languageEnglish (US)
Pages (from-to)147-152
Number of pages6
JournalNature Methods
Volume8
Issue number2
DOIs
StatePublished - Feb 2011
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Biochemistry
  • Biotechnology
  • Cell Biology

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