Scanless two-photon excitation with temporal focusing

Abstract

This Review discusses temporal focusing microscopy and its applications in neuroscience for imaging and optogenetic activation.

An azo dye for photodynamic therapy that is activated selectively by two-photon excitation

“… 6 Additionally, TP-PDT can be used with greater spatial precision owing to the quadratic dependence of TPE on the local photon intensity. 7 Hence, the production of ROS can be controlled at the confined region to the focal plane of the laser beam, enabling treatment of the target pathologic site deep inside the tissue without damaging the surrounding healthy tissue. 8 In combination with TPE, PS, which generates cytotoxic ROS via hypoxia-tolerant electron transfer type I photoreaction is beneficial.…”

Fast in vivo multiphoton light-sheet microscopy with optimal pulse frequency

“…This limit sets important constraints for investigating fast biological phenomena or for multiscale imaging [3]. Improving the speed of multiphoton microscopy is therefore an active field of research [4][5][6][7][8][9]. Among the strategies developed to improve the acquisition speed in multiphoton fluorescence imaging, light-sheet illumination exhibits distinctive advantages [4], with applications in cell biology [10], neurosciences [11], developmental biology [12], or organoid research [13].…”

“…Among the strategies developed to improve the acquisition speed in multiphoton fluorescence imaging, light-sheet illumination exhibits distinctive advantages [4], with applications in cell biology [10], neurosciences [11], developmental biology [12], or organoid research [13]. Light-sheet illumination involves reduced average power and peak intensity [4] compared to other approaches based on a collinear arrangement of illumination and detection such as multifocal point-scanning [5] or scanless wide field illumination [9]. Indeed, the parallelization of the illumination in an orthogonal geometry such as in light-sheet imaging is done along the light propagation direction, requiring a single excitation beam to excite many pixels.…”

Balancing signal and photoperturbation in multiphoton light-sheet microscopy by optimizing laser pulse frequency

“…Among the strategies developed to improve the acquisition speed in multiphoton fluorescence imaging, light-sheet illumination exhibits distinctive advantages [4], with applications in cell biology [10], neurosciences [11], developmental biology [12], or organoid research [13]. Light-sheet illumination involves reduced average power and peak intensity [4] compared to other approaches based on a collinear arrangement of illumination and detection such as multifocal point-scanning [5] or scanless wide field illumination [9]. Indeed, the parallelization of the illumination in an orthogonal geometry such as light-sheet imaging is done along the light propagation direction, requiring a single excitation beam to excite many pixels.…”