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.
Bibliographical noteFunding Information:
This work is supported by the UK Engineering Physical Sciences Research Council (EPSRC). M.A. acknowledges the support of an EPSRC-funded ‘‘Rising Star’’ Fellowship and the Scottish Universities Life Sciences Alliance (SULSA). K.D. is a Royal Society Wolfson-Merit Award holder. F.G.M. acknowledges the support of E. Killick, RS MacDonald and The ‘‘BRAINS’’ 600th anniversary appeal.
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