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Femtosecond laser pulses used as a pumping pulse in two-photon excited fluorescence microscopy were adaptively shaped by two separate self-learning schemes. First, the laser pulse was delivered through a singlemode fiber, and its pulse shape was adaptively optimized before the fiber propagation so that its fluorescent efficiency reaches the highest level. Second, the laser pulse was delivered through a microstructure fiber , and the spectrum shape of the super-continuum generated by the microstructure fiber was adaptively optimized by shaping the input pulse so that the intensity ratio of fluorescence generated from a protein mixture becomes the highest.

Highly Functional Two-Photon Excited Fluorescence Microscopy with Pulse Shaping of Femtosecond Excitation Pulses