Wide-field three-photon excitation in biological samples
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Author(s)
Fukumura, Dai
Jain, Rakesh K
Rowlands, Christopher
Park, Demian
Bruns, Oliver Thomas
Piatkevich, Kiryl
Bawendi, Moungi G
Boyden, Edward
So, Peter T. C.
Date Issued
May 2017
Journal
Light: Science & Applications
Publisher
Springer Nature
Citation
Rowlands, Christopher J et al. “Wide-Field Three-Photon Excitation in Biological Samples.” Light: Science & Applications 6, 5 (November 2016): e16255 © 2017 The Author(s)
Version
Author's final manuscript
Abstract
Three-photon wide-field depth-resolved excitation is used to overcome some of the limitations in conventional point-scanning two- and three-photon microscopy. Excitation of chromophores as diverse as channelrhodopsins and quantum dots is shown, and a penetration depth of more than 700 μm into fixed scattering brain tissue is achieved, approximately twice as deep as that achieved using two-photon wide-field excitation. Compatibility with live animal experiments is confirmed by imaging the cerebral vasculature of an anesthetized mouse; a complete focal stack was obtained without any evidence of photodamage. As an additional validation of the utility of wide-field three-photon excitation, functional excitation is demonstrated by performing three-photon optogenetic stimulation of cultured mouse hippocampal neurons expressing a channelrhodopsin; action potentials could reliably be excited without causing photodamage.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Massachusetts Institute of Technology. Department of Chemistry
Massachusetts Institute of Technology. Department of Mechanical Engineering
Massachusetts Institute of Technology. Media Laboratory
McGovern Institute for Brain Research at MIT
Massachusetts Institute of Technology. Center for Neurobiological Engineering
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DOI of Published Version
https://doi.org/10.1038/LSA.2016.255
