Improved image contrast in nonlinear light-sheet fluorescence microscopy using i²PIE Pulse compression

Nonlinear microscopy has become an invaluable tool for biological imaging, offering high-resolution visualization of biological specimens. In this manuscript, we present the application of a spectral phase measurement technique, i ² 2 PIE, to compress broad-bandwidth supercontinuum pulses for two-photon excitation fluorescence light-sheet fluorescence microscopy. The results demonstrated a significant improvement in the two-photon excitation response achieved. We also showed that the implementation of i ² 2 PIE allowed for enhanced image contrasts when compared to conventional compression techniques, with i ² 2 PIE producing an image contrast improvement over conventional methods by over 50%.