Abstract

The 3D structured illumination method is a powerful tool to improve the resolution beyond diffraction limitation in both lateral direction and axial direction [1]. However, the 3D structured excitation light, which is wide field, has no ability to confine the excitation along the axial direction. If we apply this method for deep imaging, huge background fluorescence, which sacrifices the signal to background ratio of the raw image, is generated around the surface. Temporal focusing (TF) technique provides both wide field illumination and axial excitation confinement ability for multiphoton microscopy. Previously, we combined the 2D structured illumination method with TF to remove the emission background and increase the spatial resolution in deep imaging [2]. Here we extend the idea to combine the 3D structured illumination method with TF in order to further increase the sectioning ability. To our knowledge, this is the first time to demonstrate a multiphoton 3D structured illumination microscopy. In this talk, we will give both theoretical explanation and experimental proof.

© 2015 IEEE