Abstract
Stochastic distortion of light beams in scattering samples makes in-depth photoexcitation in brain tissue a major challenge. A common solution for overcoming scattering involves adaptive pre-compensation of the unknown distortion1,2,3. However, this requires long iterative searches for sample-specific optimized corrections, which is a problem when applied to optical neurostimulation where typical timescales in the system are in the millisecond range. Thus, photoexcitation in scattering media that is independent of the properties of a specific sample would be an ideal solution. Here, we show that temporally focused two-photon excitation4 with generalized phase contrast5 enables photoexcitation of arbitrary spatial patterns within turbid tissues with remarkable robustness to scattering. We demonstrate three-dimensional confinement of tailored photoexcitation patterns >200 µm in depth, both in numerical simulations and through brain slices combined with patch-clamp recording of photoactivated channelrhodopsin-2.
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Acknowledgements
The authors thank J. Montanaro, G. Bouchery and A. Schorscher-Petcu for the preparation of fixed slices, M. Briand for her contribution to data analysis and A. Teitelboim for her help in preparing scattering liquid solutions. E.P. was supported by the Fondation pour la Recherche Médicale (FRM). A.B. was supported by the Ecole des Neurosciences de Paris (ENP). V.E. was supported by the Human Frontier Science Program (RGP0013/2010) and the Fondation pour la Recherche Médicale (FRM équipe). D.O. acknowledges support by a European Research Council starting investigator grant (no. SINSLIM 258221) and the Crown Center of Photonics. O.S. is supported by the Adams Fellowships programme of the Israel Academy of Sciences and Humanities. The Emiliani group would like to thank A. Marty and I. Llano for providing space to host the optical set-up during the renovation works in the laboratory and D. Ogden for providing scientific equipment for the electrophysiology setup.
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E.P., A.B. and B.L. performed the optical experiments. J.B. performed viral expression. E.P. built the optical set-up for electrophysiology experiments, and A.B. and B.S. performed electrophysiology recordings. B.L. and O.S. performed numerical simulations. D.O. and V.E. conceived and supervised the project, and wrote the manuscript with contributions from all co-authors.
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Papagiakoumou, E., Bègue, A., Leshem, B. et al. Functional patterned multiphoton excitation deep inside scattering tissue. Nature Photon 7, 274–278 (2013). https://doi.org/10.1038/nphoton.2013.9
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DOI: https://doi.org/10.1038/nphoton.2013.9
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