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Two-Photon Excitation Microscopy and Its Applications in Neuroscience

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Advanced Fluorescence Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1251))

Abstract

Two-photon excitation (2PE) overcomes many challenges in fluorescence microscopy. Compared to confocal microscopy, 2PE microscopy improves depth penetration, owing to the longer excitation wavelength required and to the ability to collect scattered emission photons as a useful signal. It also minimizes photodamage because lower energy photons are used and because fluorescence is confined to the geometrical focus of the laser spot. 2PE is therefore ideal for high-resolution, deep-tissue, time-lapse imaging of dynamic processes in cell biology. Here, we provide examples of important applications of 2PE for in vivo imaging of neuronal structure and signals; we also describe how it can be combined with optogenetics or photolysis of caged molecules to simultaneously probe and control neuronal activity.

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Acknowledgments

This work was supported by the Stein Oppenheimer Endowment Award and by grants from the US National Institutes of Health (5R01HD054453 from the Eunice Kennedy Shriver National Institute of Child Health and Human Development and 5RC1NS068093 from the National Institute of Neurological Disorders and Stroke).

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Authors and Affiliations

  1. Department of Neurology, David Geffen School of Medicine at UCLA, 710 Westwood Plaza, Los Angeles, CA, 90095, USA

    Ricardo Mostany, Amaya Miquelajauregui, Matthew Shtrahman & Carlos Portera-Cailliau

  2. Department of Pharmacology, Tulane University School of Medicine, 1430 Tulane Avenue, New Orleans, LA, 70112, USA

    Ricardo Mostany

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  1. Ricardo Mostany
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  2. Amaya Miquelajauregui
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  3. Matthew Shtrahman
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  4. Carlos Portera-Cailliau
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Correspondence to Carlos Portera-Cailliau .

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  1. Department of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany

    Peter J. Verveer

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Mostany, R., Miquelajauregui, A., Shtrahman, M., Portera-Cailliau, C. (2015). Two-Photon Excitation Microscopy and Its Applications in Neuroscience. In: Verveer, P. (eds) Advanced Fluorescence Microscopy. Methods in Molecular Biology, vol 1251. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2080-8_2

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