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Scanless two-photon excitation with temporal focusing
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Scanless two-photon excitation with temporal focusing
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Journal Article
Scanless two-photon excitation with temporal focusing
2020
Overview
Temporal focusing, with its ability to focus light in time, enables scanless illumination of large surface areas at the sample with micrometer axial confinement and robust propagation through scattering tissue. In conventional two-photon microscopy, widely used for the investigation of intact tissue in live animals, images are formed by point scanning of a spatially focused pulsed laser beam, resulting in limited temporal resolution of the excitation. Replacing point scanning with temporally focused widefield illumination removes this limitation and represents an important milestone in two-photon microscopy. Temporal focusing uses a diffusive or dispersive optical element placed in a plane conjugate to the objective focal plane to generate position-dependent temporal pulse broadening that enables axially confined multiphoton absorption, without the need for tight spatial focusing. Many techniques have benefitted from temporal focusing, including scanless imaging, super-resolution imaging, photolithography, uncaging of caged neurotransmitters and control of neuronal activity via optogenetics.
This Review discusses temporal focusing microscopy and its applications in neuroscience for imaging and optogenetic activation.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ Animals
/ Biomedical and Life Sciences
/ Biomedical Engineering/Biotechnology
/ Genetics
/ Image Enhancement - instrumentation
/ Imaging, Three-Dimensional - instrumentation
/ Imaging, Three-Dimensional - methods
/ Light
/ Microscopy, Fluorescence, Multiphoton - instrumentation
/ Microscopy, Fluorescence, Multiphoton - methods
/ Optics
/ Photons
/ Physics
/ Scanning
