Aberration-free three-dimensional multiphoton imaging of neuronal activity at kHz rates

Edward J. Botcherby, Christopher W. Smith, Michael M. Kohl, Delphine Débarre, Martin J. Booth, Rimas Juškaitis, Ole Paulsen, and Tony Wilson PNAS 109(8), 2919-2924 (February 2012) doi:10.1073/pnas.1111662109

Multiphoton microscopy is a powerful tool in neuroscience, promising to deliver important data on the spatiotemporal activity within individual neurons as well as in networks of neurons. A major limitation of current technologies is the relatively slow scan rates along the z direction compared to the kHz rates obtainable in the x and y directions. Here, we describe a custom-built microscope system based on an architecture that allows kHz scan rates over hundreds of microns in all three dimensions without introducing aberration. We further demonstrate how this high-speed 3D multiphoton imaging system can be used to study neuronal activity at millisecond resolution at the subcellular as well as the population level.

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