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 ca...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.展开更多
基金This work was funded by The National Key R&D Program of China(2016YFF0102000)Scientific Research and Equipment Development Project of CAS(YJKYYQ20180032 and YJKYYQ20190048)Major Innovative Research Team of Suzhou(ZXT2019007).
文摘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.
