Abstract
By integrating ultrafast laser pulse shaping into temporal focusing two-photon microscopy, a high-speed 3D imaging method is developed. This 3D imaging system requires neither laser beam steering nor sample mechanical scanning. The z scanning is achieved by shifting the temporal focal plane via applying different group velocity dispersions on the femtosecond laser spectrum in the temporal focusing two-photon microscope, and this group velocity dispersion control is done with the pulse shaping method by applying modulation functions on an acoustic optic modulator which diffracts the laser spectrum. The dependence of scanning depth on the applied electronic signals which can be tuned at kHz speed was characterized. Its high-speed 3D imaging capability was demonstrated by imaging fluorescence microspheres in a volume of 100 × 100 × 80 µm3.
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