Optogenetic manipulation of neural activity in freely moving Caenorhabditis elegans
Department of Neurobiology; Alkema Lab
Animals; Caenorhabditis elegans; *Movement; Muscle Cells; Neurons; *Optical Processes; Photobiology
Neuroscience and Neurobiology
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 ( approximately 50 frames per second) to provide high spatial resolution ( approximately 30 mum). 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.
DOI of Published Version
Nat Methods. 2011 Feb;8(2):147-52. Epub 2011 Jan 16. Link to article on publisher's site
Leifer, Andrew M.; Fang-Yen, Christopher; Gershow, Marc; Alkema, Mark J.; and Samuel, Aravinthan D. T., "Optogenetic manipulation of neural activity in freely moving Caenorhabditis elegans" (2011). Neurobiology Publications and Presentations. 3.