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Review
. 2020 Jun;17(6):571-581.
doi: 10.1038/s41592-020-0795-y. Epub 2020 Apr 13.

Scanless two-photon excitation with temporal focusing

Eirini Papagiakoumou  1 , Emiliano Ronzitti  1 , Valentina Emiliani  2
Affiliations
Review

Scanless two-photon excitation with temporal focusing

Eirini Papagiakoumou et al. Nat Methods. 2020 Jun.

Abstract

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.

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