Two-photon microscope for multisite microphotolysis of caged neurotransmitters in acute brain slices

Bradley Edward Losavio; Vijay Iyer; Peter Saggau

doi:10.1117/1.3275468

1 November 2009 Two-photon microscope for multisite microphotolysis of caged neurotransmitters in acute brain slices

Bradley Edward Losavio, Vijay Iyer, Peter Saggau

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Journal of Biomedical Optics, Vol. 14, Issue 6, 064033 (November 2009). https://doi.org/10.1117/1.3275468

Abstract

We developed a two-photon microscope optimized for physiologically manipulating single neurons through their postsynaptic receptors. The optical layout fulfills the stringent design criteria required for high-speed, high-resolution imaging in scattering brain tissue with minimal photodamage. We detail the practical compensation of spectral and temporal dispersion inherent in fast laser beam scanning with acousto-optic deflectors, as well as a set of biological protocols for visualizing nearly diffraction-limited structures and delivering physiological synaptic stimuli. The microscope clearly resolves dendritic spines and evokes electrophysiological transients in single neurons that are similar to endogenous responses. This system enables the study of multisynaptic integration and will assist our understanding of single neuron function and dendritic computation.

©(2009) Society of Photo-Optical Instrumentation Engineers (SPIE)

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Bradley Edward Losavio, Vijay Iyer, and Peter Saggau "Two-photon microscope for multisite microphotolysis of caged neurotransmitters in acute brain slices," Journal of Biomedical Optics 14(6), 064033 (1 November 2009). https://doi.org/10.1117/1.3275468

Published: 1 November 2009

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