Super-resolution imaging has revolutionized our ability to visualize biological structures at subcellular scales.However,deep-tissue super-resolution imaging remains constrained by background interference,which leads ...Super-resolution imaging has revolutionized our ability to visualize biological structures at subcellular scales.However,deep-tissue super-resolution imaging remains constrained by background interference,which leads to limited depth penetration and compromised imaging fidelity.To overcome these challenges,we propose a novel imaging system,confocal2 spinning-disk image scanning microscopy(C^(2)SD-ISM).It integrates a spinning-disk(SD)confocal microscope,which physically eliminates out-of-focus signals,forming the first confocal level.A digital micromirror device(DMD)is employed for sparse multifocal illumination,combined with a dynamic pinhole array pixel reassignment(DPA-PR)algorithm for ISM super-resolution reconstruction,forming the second confocal level.The dual confocal configuration enhances system resolution,while effectively mitigating scattering background interference.Compared to computational out-of-focus signal removal,SD preserves the original intensity distribution as the penetration depth increases,achieving an imaging depth of up to 180μm.Additionally,the DPA-PR algorithm effectively corrects Stokes shifts,optical aberrations,and other non-ideal conditions,achieving a lateral resolution of 144 nm and an axial resolution of 351 nm,and a linear correlation of up to 92%between the original confocal and the reconstructed image,thereby enabling high-fidelity super-resolution imaging.Moreover,the system's programmable illumination via the DMD allows for seamless realization with structured illumination microscopy modality,offering excellent scalability and ease of use.Altogether,these capabilities make the C^(2)SD-ISM system a versatile tool,advancing cellular imaging and tissue-scale exploration for modern bioimaging needs.展开更多
Owing to their unique pattern and abundant chemical composition, latent fingerprints （LFPs） can serve as ＂ID cards＂ and ＂information banks＂ of donors and therefore are valuable for forensic investigation, access...Owing to their unique pattern and abundant chemical composition, latent fingerprints （LFPs） can serve as ＂ID cards＂ and ＂information banks＂ of donors and therefore are valuable for forensic investigation, access control, and even medical diagnosis. LFP imaging has attracted considerable attention, and a great variety of contrast agents has been developed. In LFP imaging, background signals such as background fluorescence from the underlying surface can seriously blur the LFP images and decrease imaging sensitivity; thus, great efforts have been made to eliminate background interference. Here, we stratify the recent progress in background-free LFP imaging by making use of the difference in properties between contrast agents and background compounds. For example, near-infrared （NIR） light-activatable contrast agents can efficiently remove background signals in LFP imaging because the background compounds cannot be excited by NIR light, showing that the difference in excitation properties between contrast agents and background compounds can be employed to eliminate background interference. This review is organized around background-free LFP imaging based on the difference in optical properties between contrast agents and background compounds： （i） different excitation wavelengths, （ii） different emission wavelengths, （iii） different luminescence lifetime values, （iv） different plasmonic properties, （v） different photothermal properties, and （vi） different electrochemiluminescence properties.展开更多
基金supported by the National Key R&D Program of China(2022YFC3401100)the National Natural Science Foundation of China(62025501,92150301,62335008,62405010,and 62305004)the Postdoctoral Fellowship Program of CPSF(GZB20250669)。
文摘Super-resolution imaging has revolutionized our ability to visualize biological structures at subcellular scales.However,deep-tissue super-resolution imaging remains constrained by background interference,which leads to limited depth penetration and compromised imaging fidelity.To overcome these challenges,we propose a novel imaging system,confocal2 spinning-disk image scanning microscopy(C^(2)SD-ISM).It integrates a spinning-disk(SD)confocal microscope,which physically eliminates out-of-focus signals,forming the first confocal level.A digital micromirror device(DMD)is employed for sparse multifocal illumination,combined with a dynamic pinhole array pixel reassignment(DPA-PR)algorithm for ISM super-resolution reconstruction,forming the second confocal level.The dual confocal configuration enhances system resolution,while effectively mitigating scattering background interference.Compared to computational out-of-focus signal removal,SD preserves the original intensity distribution as the penetration depth increases,achieving an imaging depth of up to 180μm.Additionally,the DPA-PR algorithm effectively corrects Stokes shifts,optical aberrations,and other non-ideal conditions,achieving a lateral resolution of 144 nm and an axial resolution of 351 nm,and a linear correlation of up to 92%between the original confocal and the reconstructed image,thereby enabling high-fidelity super-resolution imaging.Moreover,the system's programmable illumination via the DMD allows for seamless realization with structured illumination microscopy modality,offering excellent scalability and ease of use.Altogether,these capabilities make the C^(2)SD-ISM system a versatile tool,advancing cellular imaging and tissue-scale exploration for modern bioimaging needs.
基金This work was supported by the National Natural Science Foundation of China （No. 21675120）, the National Key R＆D Program of China （Nos. 2017YFA0208000 and 2016YFF0100800）, the National Basic Research Program of China （973 Program, No. 2015CB932600） and Ten Thousand Talents Program for Young Talents.
文摘Owing to their unique pattern and abundant chemical composition, latent fingerprints （LFPs） can serve as ＂ID cards＂ and ＂information banks＂ of donors and therefore are valuable for forensic investigation, access control, and even medical diagnosis. LFP imaging has attracted considerable attention, and a great variety of contrast agents has been developed. In LFP imaging, background signals such as background fluorescence from the underlying surface can seriously blur the LFP images and decrease imaging sensitivity; thus, great efforts have been made to eliminate background interference. Here, we stratify the recent progress in background-free LFP imaging by making use of the difference in properties between contrast agents and background compounds. For example, near-infrared （NIR） light-activatable contrast agents can efficiently remove background signals in LFP imaging because the background compounds cannot be excited by NIR light, showing that the difference in excitation properties between contrast agents and background compounds can be employed to eliminate background interference. This review is organized around background-free LFP imaging based on the difference in optical properties between contrast agents and background compounds： （i） different excitation wavelengths, （ii） different emission wavelengths, （iii） different luminescence lifetime values, （iv） different plasmonic properties, （v） different photothermal properties, and （vi） different electrochemiluminescence properties.
