Two-photon structured illumination microscopy imaging using Fourier ptychography scheme

Abstract

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.