Improving resolution of second harmonic generation microscopy via scanning structured illumination

Second harmonic generation microscopy (SHGM) is a well-known technique for examining the noncentrosymmetric structures in biomedical research. However, without real-state transitions, fluorescence-based superresolution methods cannot be applied. To improve the resolution, fringe-scanning SHGM (FS-SHGM), which combines SHGM with structured illumination based on point-scanning, is introduced in this paper. The scanning path was modulated to generate illumination patterns. For the coherent parts of SHG signals, a mathematical model of image formation and reconstruction was established. Both simulations and experiments showed a resolution improvement factor of ~1.4 in the lateral and 1.56 in the axial directions for chicken tendons and mouse skin.