Thermal lensing effects in two-photon light-sheet microscopy.

ABSTRACT: In light-sheet fluorescence microscopy (LSFM), the axial resolution is governed by the illumination beam profile, motivating the development of advanced beam-shaping techniques to enhance imaging performance. Two-photon LSFM (2P-LSFM), in particular, improves the signal-to-background ratio by reducing laser scattering and distortion in biological specimens. However, we report a potentially detrimental thermal effect in 2P-LSFM: the high laser powers required for two-photon excitation induce localized heating, which alters the refractive index of the medium and effectively forms a divergent thermal lens in water. At 500 mW, the light-sheet waist broadens by 25% and shifts by 300 μm before stabilizing several seconds after the laser shutter is opened. Both experiments and simulations reveal

SUBMITTER: Hubert A 

PROVIDER: S-EPMC12904557 | biostudies-literature | 2026 Feb

REPOSITORIES: biostudies-literature