Since the 1990s,continuous technical and scientific advances have defied the diffraction limit in microscopy and enabled three-dimensional(3D)super-resolution imaging.An important milestone in this pursuit is the cohe...Since the 1990s,continuous technical and scientific advances have defied the diffraction limit in microscopy and enabled three-dimensional(3D)super-resolution imaging.An important milestone in this pursuit is the coherent utilization of two opposing objectives(4Pi geometry)and its combination with superresolution microscopy.Herein,we review the recent progress in 4Pi nanoscopy,which provides a 3D,non-invasive,diffraction-unlimited,and isotropic resolution in transparent samples.This review includes both the targeted and stochastic switching modalities of 4Pi nanoscopy.The schematics,principles,applications,and future potential of 4Pi nanoscopy are discussed in detail.展开更多
Deterministic three-dimensional(3D)super-resolution microscopy can achieve light-matter interaction in a small volume,but usually with the axial extension distinctly more elongated than the lateral one.The isoSTED met...Deterministic three-dimensional(3D)super-resolution microscopy can achieve light-matter interaction in a small volume,but usually with the axial extension distinctly more elongated than the lateral one.The isoSTED method combining two opposing objectives and multiple laser beams can offer high axial extension atλ/12 level,but at the cost of optical system complexity and inherent sidelobes.The high-order nonlinear effect by multiphoton excitation would benefit to achieve a sub-diffraction resolution as well as to suppress the sidelobes.Herein,to achieve an easyto-use,sidelobe-free deterministic 3D nanoscopy with high axial resolution,we developed a purely physical deterministic strategy(UNEx-4Pi)by fusion of ultrahighly nonlinear excitation(UNEx)of photon avalanching nanoparticles and mirror-based bifocal vector field modulation(4Pi).The theoretical studies of UNEx-4Pi concept showed that the main peak of fluorescence spot became sharper and its large sidelobe height was suppressed with the increasing optical nonlinearity.In addition,the simplicity and robustness of UNEx-4Pi system were demonstrated utilizing a mirror-assisted single-objective bifocal self-interference strategy.Experimentally,UNEx-4Pi realized an extremely constringent focal spot without sidelobes observed,achieving an axial resolution up toλ/33(26 nm)using one low-power CW beam.We also demonstrated the super-resolution ability of the UNEx-4Pi scheme to bioimaging and nuclear envelope of BSC-1 cells were stained and imaged at an axial resolution of 32 nm.The proposed UNEx-4Pi method will pave the way for achieving light-matter interaction in a highly confined space,thereby advancing cuttingedge technologies like deterministic super-resolution sensing,imaging,lithography,and data storage.展开更多
基金financially supported by the grants from National Key Research and Development Program of China (2018YFA0701400 and 2018YFE0119000)the Fundamental Research Funds for the Central Universities (2019QNA5006)+2 种基金ZJU-Sunny Photonics Innovation Center (2019-01)Zhejiang Provincial Natural Science Foundation of China (LR18H180001)startup grant from Southern University of Science and Technology
文摘Since the 1990s,continuous technical and scientific advances have defied the diffraction limit in microscopy and enabled three-dimensional(3D)super-resolution imaging.An important milestone in this pursuit is the coherent utilization of two opposing objectives(4Pi geometry)and its combination with superresolution microscopy.Herein,we review the recent progress in 4Pi nanoscopy,which provides a 3D,non-invasive,diffraction-unlimited,and isotropic resolution in transparent samples.This review includes both the targeted and stochastic switching modalities of 4Pi nanoscopy.The schematics,principles,applications,and future potential of 4Pi nanoscopy are discussed in detail.
基金supported by National Key Research and Development Program of China(2023YFF0722600)the National Natural Science Foundation of China(62335008,62122028,62105106)+3 种基金Guangdong Basic and Applied Basic Research Foundation(2023B1515040018,2022A1515011395,2024A1515012073)China Postdoctoral Science Foundation(2023T160237)the Guangdong College Student Scientific and Technological Innovation‘Climbing Program’Special Fund(pdjh2024a108)the Scientific Research Innovation Project of Graduate School of South China Normal University.
文摘Deterministic three-dimensional(3D)super-resolution microscopy can achieve light-matter interaction in a small volume,but usually with the axial extension distinctly more elongated than the lateral one.The isoSTED method combining two opposing objectives and multiple laser beams can offer high axial extension atλ/12 level,but at the cost of optical system complexity and inherent sidelobes.The high-order nonlinear effect by multiphoton excitation would benefit to achieve a sub-diffraction resolution as well as to suppress the sidelobes.Herein,to achieve an easyto-use,sidelobe-free deterministic 3D nanoscopy with high axial resolution,we developed a purely physical deterministic strategy(UNEx-4Pi)by fusion of ultrahighly nonlinear excitation(UNEx)of photon avalanching nanoparticles and mirror-based bifocal vector field modulation(4Pi).The theoretical studies of UNEx-4Pi concept showed that the main peak of fluorescence spot became sharper and its large sidelobe height was suppressed with the increasing optical nonlinearity.In addition,the simplicity and robustness of UNEx-4Pi system were demonstrated utilizing a mirror-assisted single-objective bifocal self-interference strategy.Experimentally,UNEx-4Pi realized an extremely constringent focal spot without sidelobes observed,achieving an axial resolution up toλ/33(26 nm)using one low-power CW beam.We also demonstrated the super-resolution ability of the UNEx-4Pi scheme to bioimaging and nuclear envelope of BSC-1 cells were stained and imaged at an axial resolution of 32 nm.The proposed UNEx-4Pi method will pave the way for achieving light-matter interaction in a highly confined space,thereby advancing cuttingedge technologies like deterministic super-resolution sensing,imaging,lithography,and data storage.
