Light-sheet light-field fluorescence microscopy

Light sheet fluorescence microscopy (LSFM) is a competitive 3D fluorescence imaging method for live biological specimens owing to its simultaneously fast volumetric imaging speed, high contrast, and low phototoxicity. Usually, the axial resolution is hard to be improved in an optical microscopy, especially in a LSFM with a light sheet illuminating. Here, the axial resolution is improved by using the light field imaging that can treat the light sheet with certain thickness as a volumetric imaging but not simply two-dimensional wide-field imaging. By us- on ing the light field reconstruction with backprojection (LFBP) approach and depth estimation algorithm, the axial resolution of the modified LSFM can be improved without loss of spatial resolution. In the proof-of-principle experiment, the axial resolution is improved to less than 0.5 mu m under a 6-mu m-thick light sheet illumination. With the proposed method, it provides a potential for realizing both high axial resolution and large field of view in a LSFM.

Automated summary

Light sheet fluorescence microscopy (LSFM) using light field imaging can improve axial resolution and provide high axial resolution and large field of view without losing spatial resolution.