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Fast targeted gene transfection and optogenetic modification of single neurons using femtosecond laser irradiation

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Antkowiak_srep03281.pdf (773.9Kb)
Date
21/11/2013
Author
Antkowiak, Maciej
Torres, Maria Leilani
Witts, Emily Charlotte
Miles, Gareth Brian
Dholakia, Kishan
Gunn-Moore, Frank J
Keywords
Molecular neuroscience
Cellular neuroscience
Genetic transduction
Biophotonics
RC0321 Neuroscience. Biological psychiatry. Neuropsychiatry
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Abstract
A prevailing problem in neuroscience is the fast and targeted delivery of DNA into selected neurons. The development of an appropriate methodology would enable the transfection of multiple genes into the same cell or different genes into different neighboring cells as well as rapid cell selective functionalization of neurons. Here, we show that optimized femtosecond optical transfection fulfills these requirements. We also demonstrate successful optical transfection of channelrhodopsin-2 in single selected neurons. We extend the functionality of this technique for wider uptake by neuroscientists by using fast three-dimensional laser beam steering enabling an image-guided “point-and-transfect” user-friendly transfection of selected cells. A sub-second transfection timescale per cell makes this method more rapid by at least two orders of magnitude when compared to alternative single-cell transfection techniques. This novel technology provides the ability to carry out large-scale cell selective genetic studies on neuronal ensembles and perform rapid genetic programming of neural circuits.
Citation
Antkowiak , M , Torres , M L , Witts , E C , Miles , G B , Dholakia , K & Gunn-Moore , F J 2013 , ' Fast targeted gene transfection and optogenetic modification of single neurons using femtosecond laser irradiation ' , Scientific Reports , vol. 3 , 3281 . https://doi.org/10.1038/srep03281
Publication
Scientific Reports
Status
Peer reviewed
DOI
https://doi.org/10.1038/srep03281
ISSN
2045-2322
Type
Journal article
Rights
(c) The authors. This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
Description
This work is supported by the UK Engineering Physical Sciences Research Council (EPSRC).
Collections
  • University of St Andrews Research
URI
http://hdl.handle.net/10023/4411

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