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We report on the highly nonlinear behavior observed in the central nervoussystem tissue of zebrafish (Danio rerio) when exposed to femtosecond pulses at1030 nm. At this irradiation wavelength, photo damage becomes detectable onlyafter exceeding a specific peak intensity threshold, which is independent ofthe photon flux and irradiation time, distinguishing it from irradiation atshorter wavelengths. Furthermore, we investigate and quantify the role ofexcessive heat in reducing the damage threshold, particularly duringhigh-repetition-rate operations, which are desirable for label-free andmulti-dimensional microscopy techniques. To verify our findings, we examinedcellular responses to tissue damage, including apoptosis and the recruitment ofmacrophages and fibroblasts at different time points post-irradiation. Thesefindings substantially contribute to advancing the emerging nonlinear opticalmicroscopy techniques and provide a strategy for inducing deep-tissue, preciseand localized injuries using near-infrared femtosecond laser pulses.

Highly Nonlinear Dynamics of In Vivo Deep-Tissue Interaction with Femtosecond Laser Pulses at 1030 nm