Efficient generation of high-contrast Bessel light sheets: a flexible method for improved imaging

Abstract

Light-sheet fluorescence microscopy plays a pivotal role in the field of biological 3D imaging. Among its various implementations, non-diffracting light sheets have garnered significant attention due to their remarkable ability to achieve a favorable balance between field of view and resolution. However, the presence of noticeable side-lobe effects in the non-diffracting light sheets poses challenges, including decreased contrast and an increased risk of phototoxicity. While amplitude modulation-based methods effectively suppress side-lobe influences, their transmission efficiency remains suboptimal. To address these limitations, this article introduces an approach based on phase modulation, facilitating the convenient and flexible generation of light sheets that effectively suppress side-lobe effects while maintaining high transmission efficiency. Importantly, our method enables rapid determination of optimal phase parameters, successfully suppressing the peak intensity of the first and second side lobes to levels exceeding 98% and 99%, respectively. Subsequently, experimental results substantiate the light sheet's exceptional contrast-enhancing capabilities.