Two-photon imaging using adaptive phase compensated ultrashort laser pulses
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Published on: January 28, 2019
Peng Xi1, Yair Andegeko, Dmitry Pestov
1Shanghai Jiao Tong University, Department of Biomedical Engineering, Institute for Laser Medicine and Biophotonics, Shanghai 200240 China.
Abstract:
An adaptive pulse shaper controlled by multiphoton intrapulse interference phase scanning (MIIPS) was used, together with a prism-pair, to measure and cancel high-order phase distortions introduced by a high-numerical-aperture objective and other dispersive elements of a two-photon laser-scanning microscope. The delivery of broad-bandwidth (approximately 100 nm), sub-12-fs pulses was confirmed by interferometric autocorrelation measurements at the focal plane. A comparison of two-photon imaging with transform-limited and second-order-dispersion compensated laser pulses of the same energy showed a 6-to-11-fold improvement in the two-photon excitation fluorescence signal when applied to cells and tissue, and up to a 19-fold improvement in the second harmonic generation signal from a rat tendon specimen.
