Pulse shaping for reducing photodamage in multiphoton microscopy

Dmitry Pestov; Yair Andegeko; Vadim V. Lozovoy; Marcos Dantus

doi:10.1117/12.852289

26 February 2010 Pulse shaping for reducing photodamage in multiphoton microscopy

Dmitry Pestov, Yair Andegeko, Vadim V. Lozovoy, Marcos Dantus

Proceedings Volume 7569, Multiphoton Microscopy in the Biomedical Sciences X; 756926 (2010) https://doi.org/10.1117/12.852289
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

We formulate a simple strategy for mitigation of laser-induced damage through pulse shaping and demonstrate experimentally the effect of laser pulse duration on the degree of optically induced damage for two-photon microscopy imaging. We use a broadband Ti:Sapphire laser source, aided with a shaper, and adjust both the laser pulse duration and energy to maintain constant two-photon excitation efficiency. The damage is assessed by the dynamics of two-photon excited autofluorescence intensity and sample morphology during prolonged laser exposure. We observe that for a 5-μm layer of skin tissue the damage rate is independent of the pulse shape, which suggests that the primary damage (bleaching) mechanism stems from the two-photon excitation itself. For optically thick dried blood samples, taken as another example, the data suggests that the damage is driven by one-photon absorption. In the later case, it is favorable to use shorter laser pulses to mitigate photodamage while maintaining adequate intensity of two-photon excited autofluorescence.

Citation Download Citation

Dmitry Pestov, Yair Andegeko, Vadim V. Lozovoy, and Marcos Dantus "Pulse shaping for reducing photodamage in multiphoton microscopy", Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 756926 (26 February 2010); https://doi.org/10.1117/12.852289

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