Improved image contrast in nonlinear light-sheet fluorescence microscopy using i $$^2$$ 2 PIE Pulse compression

Abstract

Abstract 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%.

Keywords

• Light-sheet fluorescence microscopy
• Nonlinear microscopy
• Biophotonics