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PMID: 18187202 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Fiber-coupled light-emitting diode for localized photostimulation of neurons expressing channelrhodopsin-2.

Journal of neuroscience methods ·Vol. 169 ·No. 1 ·2008-03-30 ·Pages 27-33

Campagnola L, Wang H, Zylka MJ

Abstract

Channelrhodopsin-2 (ChR2) is a blue-light-gated ion channel that can be used to stimulate genetically defined neurons reproducibly, rapidly and non-invasively. Existing approaches for delivering light to cells expressing ChR2 rely upon microscopes, lasers, arc lamps and shutters, all of which are relatively expensive and are not readily scalable for use on more than one brain region or animal at a time. In this paper, we describe an inexpensive method for delivering blue light locally and with millisecond precision to cells expressing ChR2. We accomplished this by coupling the light from a high-intensity blue light-emitting diode (LED; XLamp XR-E from CREE) into an optical fiber. When positioned in proximity to ChR2-expressing HEK293 cells, this fiber-coupled LED provided localized illumination of up to 32mW/mm2 and generated ChR2 photocurrents as efficiently as wide-field mercury arc lamp illumination. This fiber-coupled LED was also used to photostimulate action potentials in ChR2-expressing dorsal root ganglia (DRG) sensory neurons. LED light power and pulse frequency were controlled with an inexpensive, custom-built amplifier circuit. This scalable fiber-coupled LED system can be used to deliver light independent of the microscope objective and could, in principle, deliver light in parallel to multiple brain regions or to multiple genetically engineered animals.

MeSH Terms
Animals Cell Culture Techniques/methods Cell Line Cells, Cultured Electronics/instrumentation,methods Fiber Optic Technology/instrumentation Ganglia, Spinal/metabolism,radiation effects Humans Ion Channels/metabolism,radiation effects Light Mice Mice, Inbred C57BL Microscopy/instrumentation,methods Neurons/metabolism,radiation effects Optical Fibers Patch-Clamp Techniques/instrumentation,methods Photic Stimulation/instrumentation,methods Photochemistry/instrumentation,methods Rhodopsin/metabolism,radiation effects Sensory Rhodopsins/metabolism,radiation effects Vision, Ocular/physiology,radiation effects
Chemicals
Ion Channels Sensory Rhodopsins Rhodopsin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Campagnola Luke
Department of Cell and Molecular Physiology, 5109A NRB, CB #7545, 115 Mason Farm Road, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7545, United States.
Wang Hong
Zylka Mark J
Article Info
Journal
Journal of neuroscience methods
Abbr.
J Neurosci Methods
ISSN
0165-0270
Published
2008-03-30
Epub
2007-00-26
Pages
27-33
Language
English
Region
Netherlands
NLM ID
7905558
Subset
IM
Grants
NINDS NIH HHS · 2 T32 NS007431 · United States
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