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) |
|---|---|
| Pages (from-to) | 147-152 |
| Number of pages | 6 |
| Journal | Nature Methods |
| Volume | 8 |
| Issue number | 2 |
| DOIs | |
| State | Published - Feb 2011 |
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ASJC Scopus subject areas
- Biotechnology
- Molecular Biology
- Biochemistry
- Cell Biology
Cite this
Optogenetic manipulation of neural activity in freely moving Caenorhabditis elegans. / Leifer, Andrew M.; Fang-Yen, Christopher; Gershow, Marc; Alkema, Mark J.; Samuel, Aravinthan D T.
In: Nature Methods, Vol. 8, No. 2, 02.2011, p. 147-152.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Optogenetic manipulation of neural activity in freely moving Caenorhabditis elegans
AU - Leifer, Andrew M.
AU - Fang-Yen, Christopher
AU - Gershow, Marc
AU - Alkema, Mark J.
AU - Samuel, Aravinthan D T
PY - 2011/2
Y1 - 2011/2
N2 - 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.
AB - 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.
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U2 - 10.1038/nmeth.1554
DO - 10.1038/nmeth.1554
M3 - Article
VL - 8
SP - 147
EP - 152
JO - Nature Methods
JF - Nature Methods
SN - 1548-7091
IS - 2
ER -