Multi-scan structured illumination microscopy for rapid and efficient volumetric super-resolution imaging

Abstract

Structured illumination microscopy (SIM) is a widely adopted super-resolution imaging technique. Conventional 3D-SIM requires at least 15 exposures at each z-plane to achieve ∼2 × improved lateral and axial resolution. However, this requirement for a large number of exposures for “super-resolution” exacerbates photobleaching and slows imaging speed, thus significantly limiting its application in volumetric biological imaging. Here, we introduce multi-scan SIM (MS-SIM) that integrates a simple beam splitter for simultaneously imaging three different focal planes and a deformable mirror that enables rapid z-scanning over three contiguous sub-volumes. We demonstrate the MS-SIM system through high-quality live whole cell SIM imaging at ∼1 Hz. The high efficiency and flexibility of MS-SIM can significantly impact 3D super-resolution imaging of biological and dense colloidal systems.