Optogenetic manipulation of neural activity in freely moving Caenorhabditis elegans.
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.
Department of Physics and Center for Brain Science, Harvard University, Cambridge, Massachusetts, USA.
Andrew M
AM
Leifer
Christopher
C
Fang-Yen
Marc
M
Gershow
Mark J
MJ
Alkema
Aravinthan D T
AD
Samuel
D000818:Animals; D017173:Caenorhabditis elegans / Q000502:physiology*; D009068:Movement; D032342:Muscle Cells / Q000502:physiology; D009474:Neurons / Q000502:physiology*; D055620:Optical Phenomena; D018462:Photobiology / Q000379:methods*
Nat Methods. 2006 Oct;3(10):785-92
16990810
Int Rev Neurobiol. 2006;69:125-67
16492464
Nat Neurosci. 2005 Sep;8(9):1263-8
16116447
J Neurosci. 1985 Apr;5(4):956-64
3981252
Proc Natl Acad Sci U S A. 2006 Mar 21;103(12):4723-8
16537428
Dev Biol. 1981 Mar;82(2):358-70
7227647
Nat Methods. 2009 Dec;6(12):891-6
19898486
J Neurobiol. 2001 Jan;46(1):29-40
11108613
J Neurosci. 1996 Jun 15;16(12):4017-31
8656295
Philos Trans R Soc Lond B Biol Sci. 1976 Aug 10;275(938):327-48
8806
Curr Biol. 2005 Dec 20;15(24):2279-84
16360690
PLoS One. 2007 Mar 21;2(3):e299
17375185
Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10620-4
7938002
Neuron. 2007 May 24;54(4):535-45
17521567
Nat Methods. 2008 Oct;5(10):895-902
18794862
J Biol Chem. 2006 Mar 24;281(12):7881-9
16434400
Biol Cybern. 2008 Apr;98(4):339-51
18350313
Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):13940-5
14615590
J Neurosci. 2010 Aug 18;30(33):11151-6
20720122
Physiol Rev. 1996 Jul;76(3):687-717
8757786
J Neurosci Methods. 2010 Aug 15;191(1):90-3
20538016
Nat Neurosci. 2008 Oct;11(10):1168-76
18806786
Nature. 2010 Jan 7;463(7277):98-102
20054397
Nature. 2009 Sep 17;461(7262):407-10
19759620
J Comput Neurosci. 2008 Jun;24(3):253-76
17768672
Nature. 2007 Apr 5;446(7136):633-9
17410168
Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):12651-6
12271129