Super-resolution fluorescence microscopy plays an important role in the field of biological science with a tremendous potential, for the capability of observing living cells in real time. Numerous super-resolution methods have been developed to surpass the diffraction limit in recent years. Two-photon structured illumination microscopy (TPSIM) combines structured illumination microscopy (SIM) with two-photon excitation, providing wide field of view with deep penetration, and considerable resolution enhancement simultaneously. Here, we report a new algorithm for TPSIM termed as Fourier ptychographic (FP) technique. The result of simulation is presented to demonstrate that FP scheme is able to reduce the number of raw images with substantial resolution enhancement. The proposed method enables TPSIM to achieve live-cell imaging with fewer effective illumination patterns, shorter acquisition time, deeper imaging depth, and less phototoxicity. In addition, we show that, the number of raw images can further reduce to 4 for acceptable resolution improvement.

超分辨率荧光显微镜在生物科学领域具有重要潜力，具有实时观察活细胞的巨大潜力。近年来，已经开发了许多超分辨率方法来超过衍射极限。两光子结构照明显微镜（TPSIM）将结构照明显微镜（SIM）与两光子激发相结合，可提供宽视野和深穿透性，同时可显着提高分辨率。在这里，我们报告了一种新的TPSIM算法，称为傅里叶频谱分析（FP）技术。仿真结果表明，FP方案能够减少原始图像的数量，并且具有显着的分辨率增强。所提出的方法使TPSIM能够以更少的有效照明模式，更短的采集时间，更深的成像深度以及更少的光毒性实现活细胞成像。此外，我们表明，原始图像的数量可以进一步减少到4，以达到可接受的分辨率提高。