Sidelobe suppression in structured light sheet fluorescence microscopy by the superposition of two light sheets

Abstract

Light sheet microscopy has emerged as a powerful technique for three-dimensional and long-term vivo imaging within neuroscience and developmental biology. A light sheet illumination with structured light fields allows a better tradeoff between the field of view and axial resolution but suffers from strong side lobes. Here, we propose a method of producing structured light sheet illumination with suppressed side lobes by applying the superposition of two light sheets. The side lobe suppression results from the destructive interference between the side lobes and constructive interference between the main lobe of the two light sheets. In the proposed method, the incident light pattern in the rear pupil plane of the illumination objective is a combination of the incident light line beams required for the generation of the two interfering light sheets. We present a fast and simple calculation method to determine the incident light pattern in the rear pupil plane. Simulation results demonstrate the effectiveness of the proposed sidelobe suppression method for double-line light sheet, four-line light sheet, as well as line Bessel sheet. In particular, an 81% decrease in the relative side lobe energy can be achieved in case of double-line light sheet with an almost nonchanging propagation length. We show a way of using combined incident light patterns to generate structured light sheets with interference-resulted side lobe suppression, which is straightforward in design and with advantages of improved imaging performance.