A method to increase the imaging efficiency of tiling light sheet microscopy using scanning non-coaxial beam arrays synchronized with regional virtual confocal slits

Abstract

We present a novel method to improve the imaging efficiency of tiling light sheet microscopy. In the method, scanning non-coaxial beam arrays synchronized with regional virtual confocal slits are used to illuminate imaging plane. There are two advantages. One is the imaging efficiency increases proportional to the number of excitation beams within the non-coaxial beam array. The other is the width of the regional virtual confocal slits could be very wide without admitting off-focus fluorescence generated by the non-coaxial beam array, which makes the method easy to adopt and very robust in practice. We describe the method in detail, characterize the method via numerical simulations. The results suggest that the imaging efficiency and feasibility of the tiling light sheet microscopy could be improved significantly without affecting the 3D imaging ability by using the method. In additions, we propose several configurations to implement the method in practice.