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 language | English (US) |
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Pages (from-to) | 147-152 |
Number of pages | 6 |
Journal | Nature Methods |
Volume | 8 |
Issue number | 2 |
DOIs | |
State | Published - Feb 2011 |
Externally published | Yes |
All Science Journal Classification (ASJC) codes
- Molecular Biology
- Biochemistry
- Biotechnology
- Cell Biology