Structured Illumination Microscopy Based on Asymmetric Three-Beam Interference

Structured illumination microscopy (SIM) is a rapidly developing super-resolution technology. It has been widely used in various application fields of biomedicine due to its excellent two- and three-dimensional imaging capabilities. Furthermore, faster three-dimensional imaging methods are required to help enable more research-oriented living cell imaging. In this paper, a fast and sensitive three-dimensional structured illumination microscopy based on asymmetric three-beam interference is proposed. An innovative time-series acquisition method is employed to halve the time required to obtain each raw image. A segmented half-wave plate as a substantial linear polarization modulation method is applied to the three-dimensional SIM system for the first time. Although it needs to acquire 21 raw images instead of 15 to reconstruct one super-resolution image, the SIM setup proposed in this paper is 30% faster than the traditional spatial light modulator-SIM (SLM-SIM) in imaging each super-resolution image. The related theoretical derivation, hardware system, and verification experiment are elaborated in this paper. The stable and fast 3D super-resolution imaging method proposed in this paper is of great significance to the research of organelle interaction, intercellular communication, and other biomedical fields.