Two-photon dissociation of BeH^(+)ions is studied by detecting the fluorescence changes of Be^(+)-BeH^(+)bi-component Coulomb crystal in a linear Paul trap.BeH^(+)ions generated by an exothermic reaction between elect...Two-photon dissociation of BeH^(+)ions is studied by detecting the fluorescence changes of Be^(+)-BeH^(+)bi-component Coulomb crystal in a linear Paul trap.BeH^(+)ions generated by an exothermic reaction between electronically excited Be^(+)ions and residual H_(2) in the vacuum chamber are photon-dissociated with two photons scanning over the range of 201 nm to 208 nm.Our experiment provides a novel method to maintain the number of Be^(+)ions stable in a Coulomb crystal with a middle ultraviolet band dissociation laser.This two-photon dissociation method extends the wavelength range of the dissociation laser for BeH^(+)compared to the one-photon dissociation,and the method can be utilized to all alkaline earth atomic ions which require suppression of the reaction with residual H_(2) gas in vacuum.展开更多
Background:Glioblastoma(GBM)is a highly aggressive brain tumor characterized by aberrant angiogenesis and an immunosuppressive microenvironment.Pericytes are aberrantly recruited but their spatiotemporal roles and mol...Background:Glioblastoma(GBM)is a highly aggressive brain tumor characterized by aberrant angiogenesis and an immunosuppressive microenvironment.Pericytes are aberrantly recruited but their spatiotemporal roles and molecular changes remain unclear.This study investigated platelet-derived growth factor receptor beta-positive(Pdgfrb+)pericyte dynamics and reprogramming in GBM vasculature.Methods:We generated GL261-Luc and GL261-CFP glioblastoma cells via lentiviral transduction and established two transgenic models.(1)For pericyte labeling,Ai14 reporter mice was crossed with PDGFRβ-P2A-CreERT2mice for td Tomato-specific lineage tracing(PT mice).(2)For conditional ablation,we generated inducible Pdgfrb-expressing cell ablation models(PT mice was crossed with ROSA-DTA mice).An intravital imaging platform(FITC-dextran/CFP/td Tomato+two-photon microscopy)tracked pericytes,vessels,and tumor cells,while FACSsorted Pdgfrb+cells from GBM and normal brain were analyzed by LC-MS/MS proteomics.Results:Cre-mediated ablation of Pdgfrb-expressing cells revealed stage-dependent effects on GBM growth:early ablation inhibited progression while late ablation promoted it.Pericytes undergo dual spatial reorganization in GBM:regional enrichment with pre-sprouting accumulation at the tumor-brain interface,and focal positioning with preferential localization at vascular branch points.Concurrently,GBM vasculature displayed simplified branching,dilation,and pericyte remodeling(shorter processes,higher density).Proteomics revealed 1426 altered proteins,with upregulated proliferation pathways(e.g.,matrix metallopeptidase 14[Mmp14],lysyl oxidase like 2[Loxl2])and downregulated homeostasis functions(e.g.,transforming growth factor beta 1[Tgfb1]),validated by scRNA-seq in human GBM.Conclusions:This study demonstrates that during early GBM progression,pericytes actively drive tumor angiogenesis through molecular reprogramming toward proliferative and pro-angiogenic phenotypes,with the integrated imaging-proteomics framework revealing potential therapeutic targets for disrupting pericyte-mediated vascular remodeling.展开更多
Hypochlorous acid(HClO)is a critical biomolecule in living organisms,playing an essential role in numerous physiological or pathological processes.Abnormal levels of HClO in the body may lead to a series of diseases,f...Hypochlorous acid(HClO)is a critical biomolecule in living organisms,playing an essential role in numerous physiological or pathological processes.Abnormal levels of HClO in the body may lead to a series of diseases,for instance,inflammation and cancer.Thus,accurate measurement of HClO levels should be more beneficial for understanding its role in diseases and gaining a deeper insight into the pathogenesis of diseases.In this work,we designed a near-infrared two-photon fluorescent probe(HDM-Cl-HClO)for detecting fluctuations in HClO levels in inflammatory and tumor-bearing mice.Notably,the probe can respond to HClO within 5 s and trigger a brilliant red fluorescence at 660 nm.It exhibits high specificity and sensitivity for HClO.The superior spectral capability of the probe has enabled the detection of HClO levels in cells and zebrafish,as well as achieved the detection of HClO in inflammatory and tumor mice.This work not only provides a novel strategy and tool for HClO imaging in living systems,but also holds great potential for the diagnosis of inflammation and cancer.展开更多
Four benzocoumarin-based two-photon fluorescent probes(BH1-BH4)are proposed for ra-tiometric detection of hypochlorous acid(HClO)and their two-photon sensing perfor-mance are evaluated by means of time-dependent densi...Four benzocoumarin-based two-photon fluorescent probes(BH1-BH4)are proposed for ra-tiometric detection of hypochlorous acid(HClO)and their two-photon sensing perfor-mance are evaluated by means of time-dependent density func-tional theory and quadratic re-sponse theory.The effects of benzene-fused position on Stokes shift,fluorescence quantum yield and two-photon absorption are discussed comprehensively.The results show that fusing a benzene ring in coumarin can enhance Stokes shift efficiently.The benzene-fused position has important effects on these photophysical properties.The benzo[g]coumarins(BH1)and benzo[f]coumarins(BH2)derivatives have larger Stokes shifts in comparison with benzo[h]coumarins(BH3)and dihydrophenazine(BH4)derivatives.The two-photon absorp-tion of benzo[f]coumarins(BH2)derivative is much smaller than those of other benzo-coumarins derivatives.The large Stokes shift and increased two-photon action cross section can be achieved simultaneously in the dihydrophenazine(BH4)derivative.Therefore,the de-signed BH4 is expected to have superior performance for the ratiometric detection of HClO.To explore the reasons behind these effects,the intramolecular charge transfer degrees are il-lustrated quantitatively by plotting the hole-electron isosurface map,and the relation be-tween charge transfer and Stokes shift is revealed.A two-state model analysis is employed to understand two-photon absorption ability.Moreover,the fluorescence near-quenching mecha-nism of the product molecules B1 and B3 is explained by analyzing reorganization energy and Huang-Rhys factor,as well as related normal mode.Our research could contribute to the effi-cient design of ratiometric two-photon fluorescent probes with large Stokes shift and signifi-cant two-photon action cross section.展开更多
The complexity of cancer therapy has led to the emergence of combination therapy as a promising approach to enhance treatment efficacy and safety.The integration of glutathione(GSH)-activatable two-photon photodynamic...The complexity of cancer therapy has led to the emergence of combination therapy as a promising approach to enhance treatment efficacy and safety.The integration of glutathione(GSH)-activatable two-photon photodynamic therapy(TP-PDT)and chemodynamic therapy(CDT)offers the possibility to advance precision and efficacy in anti-cancer treatments.In this study,a GSH-activatable photosensitizer(PS),namely copper-elsinochrome(CuEC),is synthesized and utilized for combination second nearinfrared(NIR-II)TP-PDT/CDT.The Cu^(2+)acts as a“lock”,suppressing the fluorescence and^(1)O_(2)generation ability of EC in a normal physiological environment(“OFF”state).However,the overexpressed GSH in the tumor microenvironment acts as the“key”,resulting in the release of EC(“ON”state)and Cu^(+)(reduced by GSH).The released EC can be utilized for fluorescence imaging and TP-PDT under NIR-II(λ=1000 nm)two-photon excitation,while Cu+can generate highly toxic hydroxyl radicals(•OH)via Fenton-like reaction for CDT.Additionally,this process consumes GSH and diminishes the tumor’s antioxidant capacity,thereby augmenting the efficacy of combination therapy.The CuEC achieves significant tumor cell ablation in both 2D monolayer cells and 3D multicellular tumor spheres through the combination of NIR-II TP-PDT and CDT.展开更多
Optical imaging systems have greatly extended human visual capabilities,enabling the observation and understanding of diverse phenomena.Imaging technologies span a broad spectrum of wavelengths from x-ray to radio fre...Optical imaging systems have greatly extended human visual capabilities,enabling the observation and understanding of diverse phenomena.Imaging technologies span a broad spectrum of wavelengths from x-ray to radio frequencies and impact research activities and our daily lives.Traditional glass lenses are fabricated through a series of complex processes,while polymers offer versatility and ease of production.However,modern applications often require complex lens assemblies,driving the need for miniaturization and advanced designs with micro-and nanoscale features to surpass the capabilities of traditional fabrication methods.Three-dimensional(3D)printing,or additive manufacturing,presents a solution to these challenges with benefits of rapid prototyping,customized geometries,and efficient production,particularly suited for miniaturized optical imaging devices.Various 3D printing methods have demonstrated advantages over traditional counterparts,yet challenges remain in achieving nanoscale resolutions.Two-photon polymerization lithography(TPL),a nanoscale 3D printing technique,enables the fabrication of intricate structures beyond the optical diffraction limit via the nonlinear process of two-photon absorption within liquid resin.It offers unprecedented abilities,e.g.alignment-free fabrication,micro-and nanoscale capabilities,and rapid prototyping of almost arbitrary complex 3D nanostructures.In this review,we emphasize the importance of the criteria for optical performance evaluation of imaging devices,discuss material properties relevant to TPL,fabrication techniques,and highlight the application of TPL in optical imaging.As the first panoramic review on this topic,it will equip researchers with foundational knowledge and recent advancements of TPL for imaging optics,promoting a deeper understanding of the field.By leveraging on its high-resolution capability,extensive material range,and true 3D processing,alongside advances in materials,fabrication,and design,we envisage disruptive solutions to current challenges and a promising incorporation of TPL in future optical imaging applications.展开更多
Over the past decade,a growing number of studies have reported transcription factor-based in situ reprogramming that can directly conve rt endogenous glial cells into functional neurons as an alternative approach for ...Over the past decade,a growing number of studies have reported transcription factor-based in situ reprogramming that can directly conve rt endogenous glial cells into functional neurons as an alternative approach for n euro regeneration in the adult mammalian central ne rvous system.Howeve r,many questions remain regarding how a terminally differentiated glial cell can transform into a delicate neuron that forms part of the intricate brain circuitry.In addition,concerns have recently been raised around the absence of astrocyte-to-neuron conversion in astrocytic lineage-tra cing mice.In this study,we employed repetitive two-photon imaging to continuously capture the in situ astrocyte-to-neuron conversion process following ecto pic expression of the neural transcription factor NeuroD1 in both prolife rating reactive astrocytes and lineage-tra ced astrocytes in the mouse cortex.Time-lapse imaging over several wee ks revealed the ste p-by-step transition from a typical astrocyte with numero us short,tapered branches to a typical neuro n with a few long neurites and dynamic growth cones that actively explored the local environment.In addition,these lineage-converting cells were able to migrate ra dially or to ngentially to relocate to suitable positions.Furthermore,two-photon Ca2+imaging and patch-clamp recordings confirmed that the newly generated neuro ns exhibited synchronous calcium signals,repetitive action potentials,and spontaneous synaptic responses,suggesting that they had made functional synaptic connections within local neural circuits.In conclusion,we directly visualized the step-by-step lineage conversion process from astrocytes to functional neurons in vivo and unambiguously demonstrated that adult mammalian brains are highly plastic with respect to their potential for neuro regeneration and neural circuit reconstruction.展开更多
The realization of real-time thermal feedback for monitoring photothermal therapy(PTT)under near-infrared(NIR)light irradiation is of great interest and challenge for antitumor therapy.Herein,by assembling highly effi...The realization of real-time thermal feedback for monitoring photothermal therapy(PTT)under near-infrared(NIR)light irradiation is of great interest and challenge for antitumor therapy.Herein,by assembling highly efficient photothermal conversion gold nanorods and a temperature-responsive probe((E)-4-(4-(diethylamino)styryl)-1-methylpyridin-1-ium,PyS)within MOF-199,an intelligent nanoplatform(AMPP)was fabricated for simultaneous chemodynamic therapy and NIR light-induced temperature-feedback PTT.The fluorescence intensity and temperature of the PyS probe are linearly related due to the restriction of the rotation of the characteristic monomethine bridge.Moreover,the copper ions resulting from the degradation of MOF-199 in an acidic microenvironment can convert H_(2)O_(2)into•OH,resulting in tumor ablation through a Fenton-like reaction,and this process can be accelerated by increasing the temperature.This study establishes a feasible platform for fabricating highly sensitive temperature sensors for efficient temperature-feedback PTT.展开更多
To overcome the conflict between the long-wavelength excitation and high singlet oxygen quantum yield of photosensitizers,we conjugated a two-photon fluorophore,tetrahydroquinoxaline coumarin(TQ),and an efficient phot...To overcome the conflict between the long-wavelength excitation and high singlet oxygen quantum yield of photosensitizers,we conjugated a two-photon fluorophore,tetrahydroquinoxaline coumarin(TQ),and an efficient photodynamic therapeutic agent,benzo[a]phenothiazinium(NBS-NH_(2)),through a hexamethylene linker to build a two-photon photosensitizer,TQ-NBS.In TQ-NBS,TQ served as an energy donor and NBS-NH_(2) acted as an energy acceptor;and TQ-NBS was a F?rster resonance energy transfer(FRET)cassette with a 92.8%efficiency.The large two-photon absorption cross-section of TQ allowed photosensitizer TQ-NBS to work in a 900 nm two-photon excitation(TPE)mode,which greatly benefited the deep tissue penetration in PDT treatment.Meanwhile,the excellent phototoxicity and near-infrared fluorescence of NBS-NH2was kept in TQ-NBS under a TPE mode via a FRET process.Photosensitizer TQ-NBS exhibited a high phototoxic efficacy in living cells and tumor-bearing mice.展开更多
Carbazole-core multi-branched chromophores 9-ethyl- 3, 6-bis （ 2- { 4- [ 5- （4-tert-butyl-phenyl） - [ 1, 3, 4 ] oxadiazol-2-yl ] - phenyl }-vinyl） -carbazole（3） and 9-ethyl-3-（ 2- {4-[ 5-（4-tert-butyl- phenyl...Carbazole-core multi-branched chromophores 9-ethyl- 3, 6-bis （ 2- { 4- [ 5- （4-tert-butyl-phenyl） - [ 1, 3, 4 ] oxadiazol-2-yl ] - phenyl }-vinyl） -carbazole（3） and 9-ethyl-3-（ 2- {4-[ 5-（4-tert-butyl- phenyl） -[ 1, 3, 4 ] oxadiazol-2-yl ] -phenyl }-vinyl ） -carbazole （ 2 ） are synthesized through Wittig reaction and characterized by nuclear magnetic resonance（NMR）and infrared（IR）. The two- photon absorption properties of chromophores are investigated. These chromophores exhibit large two-photon absorption crosssections and strong blue two-photon excited fluorescence. The cooperative enhancement of two-photon absorption（TPA） in the multi-branched structures is observed. This enhancement is partly attributed to the electronic coupling between the branches. The electronic push-pull structures in the arm and their cooperative effects help the extended charge transfer for TPA.展开更多
The wavelength dependence of photoelectron angular distributions （PADs） of two-photon detachment of Cu^- has been directly studied by using the photoelectron map imaging. Results show that for the laser field intens...The wavelength dependence of photoelectron angular distributions （PADs） of two-photon detachment of Cu^- has been directly studied by using the photoelectron map imaging. Results show that for the laser field intensity of 6.0×10^10W/cm^2, PADs exhibit dramatic change with the external field wavelength. Comparison between the experimental observation and the lowest-order perturbation theory prediction indicates that the pattern of PADs can be explained by the interference of the s and d partial waves in the final state. Relative contri- butions of s and d partial waves in the two-photon detachment at different laser wavelengths are obtained.展开更多
Backgroud:Thoracic Trauma and Limb Fractures Are the Two most Common Injuries in Multiple Trauma.However,there Is Still a Lack of Mouse Models of Trauma Combining Tibial Shaft Fracture(TSF)and Thoracic Trauma.In this ...Backgroud:Thoracic Trauma and Limb Fractures Are the Two most Common Injuries in Multiple Trauma.However,there Is Still a Lack of Mouse Models of Trauma Combining Tibial Shaft Fracture(TSF)and Thoracic Trauma.In this Study,we Attempted to Develop a Novel Mouse Model of TSF Combined with Blunt Chest Trauma(BCT).Methods:A total of 84 C57BL/6J male mice were used as the multiple trauma model.BCT was induced by hitting the chests of mice with heavy objects,and TSF was in-duced by hitting the tibia of mice with heavy objects after intramedullary fixation.Serum specimens of mice were received by cardiac puncture at defined time points of 0,6,12,24,48,and 72 h.Results:Body weight and body temperature tended to decrease within 24 h after mul-tiple trauma.Hemoglobin analyses revealed a decrease during the first 24 h after mul-tiple trauma.Some animals died by cardiac puncture immediately after chest trauma.These animals exhibited the most severe pulmonary contusion and hemorrhage.The level of lung damage varied in diverse mice but was apparent in all animals.Classic he-matoxylin and eosin(H&E)-stained paraffin pulmonary sections of mice with multiple trauma displayed hemorrhage and an immunoinflammatory reaction.Bronchoalveolar lavage fluid(BALF)and serum samples of mice with multiple trauma showed an upreg-ulation of interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-1α(TNF-1α)compared with the control group.Microimaging confirmed the presence of a tibia fracture and pulmonary contusion.Conclusions:The novel mouse multiple trauma model established in this study is a common trauma model that shows similar pathological mechanisms and imaging characteristics in patients with multiple injuries.This study is useful for determining whether blockade or intervention of the cytokine response is beneficial for the treat-ment of patients with multiple trauma.Further research is needed in the future.展开更多
Laser processing technologies enable the precise fabrication of arbitrary structures and devices with broad applications in micro-optics,micro-mechanics,and biomedicine.However,its adoption is limited by the large siz...Laser processing technologies enable the precise fabrication of arbitrary structures and devices with broad applications in micro-optics,micro-mechanics,and biomedicine.However,its adoption is limited by the large size,complexity,high cost,and low flexibility of optical systems.Metasurfaces enable precise multidimensional control of light fields,aligning well with the development trend toward compact,high-performance optical systems.Here,we review several recent studies on the application of metasurfaces in laser processing technologies,including 3D nanolithography,direct laser writing,and laser cutting.Metasurfaces provide an integrated operational platform with exceptional performance,poised to disrupt conventional laser processing workflows.This combination presents significant cost efficiency and substantial development potential,with promising applications in areas such as imaging,optical storage,advanced sensing,and space on-orbit manufacturing.展开更多
Compound eyes(CEs)that feature ultra-compact structures and extraordinary versatility have revealed great potential for cutting-edge applications.However,the optoelectronic integration of CEs with available photodetec...Compound eyes(CEs)that feature ultra-compact structures and extraordinary versatility have revealed great potential for cutting-edge applications.However,the optoelectronic integration of CEs with available photodetectors is still challenging because the planar charge-coupled device(CCD)/complementary metal oxide semiconductor(CMOS)detector cannot match the spatially distributed images formed by CE ommatidia.To reach this end,we report here the optoelectronic integration of CEs by manufacturing 3D nonuniform ommatidia for developing an ultra-compact on-chip camera.As a proof-of-concept,we fabricated microscale CEs with uniform and nonuniform ommatidia through femtosecond laser two-photon photopolymerization,and compared their focusing/imaging performance both theoretically and experimentally.By engineering the surface profiles of the ommatidia at different positions of the CE,the images formed by all the ommatidia can be tuned on a plane.In this way,the nonuniform CE can be directly integrated with a commercial CMOS photodetector,forming an ultra-compact CE camera.Additionally,we further combine the CE camera with a microfluidic chip,which can further serve as an on-chip microscopic monitoring system.We anticipate that such an ultra-compact CE camera may find broad applications in microfluidics,robotics,and micro-optics.展开更多
Alzheimer's disease(AD)is a neurodegenerative disease characterized by a progressive decline in cognitive functions.Given that AD undermines the quality of life for millions and has an extended asymptomatic period...Alzheimer's disease(AD)is a neurodegenerative disease characterized by a progressive decline in cognitive functions.Given that AD undermines the quality of life for millions and has an extended asymptomatic period,exploring the full AD pathogenesis and seeking the optimal therapeutic solution have become critical and imperative.This allows researchers to intervene,delay,and potentially prevent AD progression.Several clinical imaging methods are utilized routinely to diagnose and monitor AD,such as magnetic resonance imaging(MRI),functional magnetic resonance imaging(fMRI),positron emission tomography(PET),and single photon emission computed tomography(SPECT).Nevertheless,due to their intrinsic drawbacks and restrictions,such as radiation concerns,high cost,long acquisition time,and low spatial resolution,their applications in AD research are limited,especially at the cellular and molecular levels.In contrast,optical microscopic imaging methods overcome these limitations,offering researchers a variety of approaches with distinct advantages to explore AD pathology on diverse models.In this review,we provide a comprehensive overview of commonly utilized optical microscopic imaging techniques in AD research and introduce their contributions to image amyloid beta(Aβ)species.These techniques include fluorescence microscopy(FM),confocal microscopy(CM),two-photon fluorescence microscopy(TPFM),super-resolution microscopy(SRM),expansion microscopy(ExM),and light-sheet fluorescence microscopy(LSFM).In addition,we introduce some related topics,such as the development of near-infrared(NIR)Aβprobes,the Aβplaque hypothesis,and Aβoligomer hypothesis,and the roles of microglia and astrocytes in AD progression.We believe optical microscopic imaging methods continue to play an indispensable role in deciphering the full pathogenesis of AD and advancing therapeutic strategies.展开更多
Highly programmable shape morphing of 4D-printed micro/nanostructures is urgently desired for applications in robotics and intelligent systems.However,due to the lack of autonomous holistic strategies throughout the t...Highly programmable shape morphing of 4D-printed micro/nanostructures is urgently desired for applications in robotics and intelligent systems.However,due to the lack of autonomous holistic strategies throughout the target shape input,optimal material distribution generation,and fabrication program output,4D nanoprinting that permits arbitrary shape morphing remains a challenging task for manual design.In this study,we report an autonomous inverse encoding strategy to decipher the genetic code for material property distributions that can guide the encoded modeling toward arbitrarily pre-programmed 4D shape morphing.By tuning the laser power of each voxel at the nanoscale,the genetic code can be spatially programmed and controllable shape morphing can be realized through the inverse encoding process.Using this strategy,the 4D-printed structures can be designed and accurately shift to the target morphing of arbitrarily hand-drawn lines under stimulation.Furthermore,as a proof-of-concept,a flexible fiber micromanipulator that can approach the target region through pre-programmed shape morphing is autonomously inversely encoded according to the localized spatial environment.This strategy may contribute to the modeling and arbitrary shape morphing of micro/nanostructures fabricated via 4D nanoprinting,leading to cutting-edge applications in microfluidics,micro-robotics,minimally invasive robotic surgery,and tissue engineering.展开更多
Three-dimensional(3D)nanoprinting via two-photon polymerization offers unparalleled design flexibility and precision,thereby enabling rapid prototyping of advanced micro-optical elements and systems that have found im...Three-dimensional(3D)nanoprinting via two-photon polymerization offers unparalleled design flexibility and precision,thereby enabling rapid prototyping of advanced micro-optical elements and systems that have found important applications in endomicroscopy and biomedical imaging.The potential of this versatile tool for monolithic manufacturing of dynamic micro-opto-electro-mechanical systems(MOEMSs),however,has not yet been sufficiently explored.This work introduces a 3D-nanoprinted lens actuator with a large optical aperture,optimized for remote focusing in miniaturized imaging systems.The device integrates orthoplanar linear motion springs,a self-aligned sintered micro-magnet,and a monolithic lens,actuated by dual microcoils for uniaxial motion.The use of 3D nanoprinting allows complete design freedom for the integrated optical lens,whereas the monolithic fabrication ensures inherent alignment of the lens with the mechanical elements.With a lens diameter of 1.4 mm and a compact footprint of 5.74 mm,it achieves high mechanical robustness at resonant frequencies exceeding 300 Hz while still providing a large displacement range of 200μm(±100μm).A comprehensive analysis of optical and mechanical performance,including the effects of coil temperature and polymer viscoelasticity,demonstrates its advantages over conventional micro-electro-mechanical system actuators,showcasing its potential for next-generation imaging applications.展开更多
基金Project supported by the National Key Research and Development Program of China(Grant No.2021YFA1402103)the National Natural Science Foundation of China(Grant No.12393825)。
文摘Two-photon dissociation of BeH^(+)ions is studied by detecting the fluorescence changes of Be^(+)-BeH^(+)bi-component Coulomb crystal in a linear Paul trap.BeH^(+)ions generated by an exothermic reaction between electronically excited Be^(+)ions and residual H_(2) in the vacuum chamber are photon-dissociated with two photons scanning over the range of 201 nm to 208 nm.Our experiment provides a novel method to maintain the number of Be^(+)ions stable in a Coulomb crystal with a middle ultraviolet band dissociation laser.This two-photon dissociation method extends the wavelength range of the dissociation laser for BeH^(+)compared to the one-photon dissociation,and the method can be utilized to all alkaline earth atomic ions which require suppression of the reaction with residual H_(2) gas in vacuum.
基金The National Key Research and Development Program of China,Grant/Award Number:2022YFF0710700Nonprofit Central Research Institute Fund of Chinese Academy of Medical Sciences,Grant/Award Number:2023-PT180-01。
文摘Background:Glioblastoma(GBM)is a highly aggressive brain tumor characterized by aberrant angiogenesis and an immunosuppressive microenvironment.Pericytes are aberrantly recruited but their spatiotemporal roles and molecular changes remain unclear.This study investigated platelet-derived growth factor receptor beta-positive(Pdgfrb+)pericyte dynamics and reprogramming in GBM vasculature.Methods:We generated GL261-Luc and GL261-CFP glioblastoma cells via lentiviral transduction and established two transgenic models.(1)For pericyte labeling,Ai14 reporter mice was crossed with PDGFRβ-P2A-CreERT2mice for td Tomato-specific lineage tracing(PT mice).(2)For conditional ablation,we generated inducible Pdgfrb-expressing cell ablation models(PT mice was crossed with ROSA-DTA mice).An intravital imaging platform(FITC-dextran/CFP/td Tomato+two-photon microscopy)tracked pericytes,vessels,and tumor cells,while FACSsorted Pdgfrb+cells from GBM and normal brain were analyzed by LC-MS/MS proteomics.Results:Cre-mediated ablation of Pdgfrb-expressing cells revealed stage-dependent effects on GBM growth:early ablation inhibited progression while late ablation promoted it.Pericytes undergo dual spatial reorganization in GBM:regional enrichment with pre-sprouting accumulation at the tumor-brain interface,and focal positioning with preferential localization at vascular branch points.Concurrently,GBM vasculature displayed simplified branching,dilation,and pericyte remodeling(shorter processes,higher density).Proteomics revealed 1426 altered proteins,with upregulated proliferation pathways(e.g.,matrix metallopeptidase 14[Mmp14],lysyl oxidase like 2[Loxl2])and downregulated homeostasis functions(e.g.,transforming growth factor beta 1[Tgfb1]),validated by scRNA-seq in human GBM.Conclusions:This study demonstrates that during early GBM progression,pericytes actively drive tumor angiogenesis through molecular reprogramming toward proliferative and pro-angiogenic phenotypes,with the integrated imaging-proteomics framework revealing potential therapeutic targets for disrupting pericyte-mediated vascular remodeling.
基金National Natural Science Foundation of China(No.22264013)Hainan Province Clinical Medical Center(No.2021)Hainan Province Science and Technology Special Fund(No.ZDYF2024SHFZ104).
文摘Hypochlorous acid(HClO)is a critical biomolecule in living organisms,playing an essential role in numerous physiological or pathological processes.Abnormal levels of HClO in the body may lead to a series of diseases,for instance,inflammation and cancer.Thus,accurate measurement of HClO levels should be more beneficial for understanding its role in diseases and gaining a deeper insight into the pathogenesis of diseases.In this work,we designed a near-infrared two-photon fluorescent probe(HDM-Cl-HClO)for detecting fluctuations in HClO levels in inflammatory and tumor-bearing mice.Notably,the probe can respond to HClO within 5 s and trigger a brilliant red fluorescence at 660 nm.It exhibits high specificity and sensitivity for HClO.The superior spectral capability of the probe has enabled the detection of HClO levels in cells and zebrafish,as well as achieved the detection of HClO in inflammatory and tumor mice.This work not only provides a novel strategy and tool for HClO imaging in living systems,but also holds great potential for the diagnosis of inflammation and cancer.
基金supported by the Shandong Provincial Natural Science Foundation,China(No.ZR2020MA078)。
文摘Four benzocoumarin-based two-photon fluorescent probes(BH1-BH4)are proposed for ra-tiometric detection of hypochlorous acid(HClO)and their two-photon sensing perfor-mance are evaluated by means of time-dependent density func-tional theory and quadratic re-sponse theory.The effects of benzene-fused position on Stokes shift,fluorescence quantum yield and two-photon absorption are discussed comprehensively.The results show that fusing a benzene ring in coumarin can enhance Stokes shift efficiently.The benzene-fused position has important effects on these photophysical properties.The benzo[g]coumarins(BH1)and benzo[f]coumarins(BH2)derivatives have larger Stokes shifts in comparison with benzo[h]coumarins(BH3)and dihydrophenazine(BH4)derivatives.The two-photon absorp-tion of benzo[f]coumarins(BH2)derivative is much smaller than those of other benzo-coumarins derivatives.The large Stokes shift and increased two-photon action cross section can be achieved simultaneously in the dihydrophenazine(BH4)derivative.Therefore,the de-signed BH4 is expected to have superior performance for the ratiometric detection of HClO.To explore the reasons behind these effects,the intramolecular charge transfer degrees are il-lustrated quantitatively by plotting the hole-electron isosurface map,and the relation be-tween charge transfer and Stokes shift is revealed.A two-state model analysis is employed to understand two-photon absorption ability.Moreover,the fluorescence near-quenching mecha-nism of the product molecules B1 and B3 is explained by analyzing reorganization energy and Huang-Rhys factor,as well as related normal mode.Our research could contribute to the effi-cient design of ratiometric two-photon fluorescent probes with large Stokes shift and signifi-cant two-photon action cross section.
基金supported by the project of the National Key Research and Development Program of China(No.2022YFA1207600)the National Natural Science Foundation of China(Nos.62005294,62375272)TIPC Director’s Fund.
文摘The complexity of cancer therapy has led to the emergence of combination therapy as a promising approach to enhance treatment efficacy and safety.The integration of glutathione(GSH)-activatable two-photon photodynamic therapy(TP-PDT)and chemodynamic therapy(CDT)offers the possibility to advance precision and efficacy in anti-cancer treatments.In this study,a GSH-activatable photosensitizer(PS),namely copper-elsinochrome(CuEC),is synthesized and utilized for combination second nearinfrared(NIR-II)TP-PDT/CDT.The Cu^(2+)acts as a“lock”,suppressing the fluorescence and^(1)O_(2)generation ability of EC in a normal physiological environment(“OFF”state).However,the overexpressed GSH in the tumor microenvironment acts as the“key”,resulting in the release of EC(“ON”state)and Cu^(+)(reduced by GSH).The released EC can be utilized for fluorescence imaging and TP-PDT under NIR-II(λ=1000 nm)two-photon excitation,while Cu+can generate highly toxic hydroxyl radicals(•OH)via Fenton-like reaction for CDT.Additionally,this process consumes GSH and diminishes the tumor’s antioxidant capacity,thereby augmenting the efficacy of combination therapy.The CuEC achieves significant tumor cell ablation in both 2D monolayer cells and 3D multicellular tumor spheres through the combination of NIR-II TP-PDT and CDT.
基金support from the National Research Foundation (NRF) Singapore, under its Competitive Research Programme Award NRF-CRP20-20170004 and NRF Investigatorship Award NRF-NRFI06-20200005MTC Programmatic Grant M21J9b0085, as well as the Lite-On Project RS-INDUS-00090+5 种基金support from Australian Research Council (DE220101085, DP220102152)grants from German Research Foundation (SCHM2655/15-1, SCHM2655/21-1)Lee-Lucas Chair in Physics and funding by the Australian Research Council DP220102152financial support from the National Natural Science Foundation of China (Grant No. 62275078)Natural Science Foundation of Hunan Province of China (Grant No. 2022JJ20020)Shenzhen Science and Technology Program (Grant No. JCYJ20220530160405013)
文摘Optical imaging systems have greatly extended human visual capabilities,enabling the observation and understanding of diverse phenomena.Imaging technologies span a broad spectrum of wavelengths from x-ray to radio frequencies and impact research activities and our daily lives.Traditional glass lenses are fabricated through a series of complex processes,while polymers offer versatility and ease of production.However,modern applications often require complex lens assemblies,driving the need for miniaturization and advanced designs with micro-and nanoscale features to surpass the capabilities of traditional fabrication methods.Three-dimensional(3D)printing,or additive manufacturing,presents a solution to these challenges with benefits of rapid prototyping,customized geometries,and efficient production,particularly suited for miniaturized optical imaging devices.Various 3D printing methods have demonstrated advantages over traditional counterparts,yet challenges remain in achieving nanoscale resolutions.Two-photon polymerization lithography(TPL),a nanoscale 3D printing technique,enables the fabrication of intricate structures beyond the optical diffraction limit via the nonlinear process of two-photon absorption within liquid resin.It offers unprecedented abilities,e.g.alignment-free fabrication,micro-and nanoscale capabilities,and rapid prototyping of almost arbitrary complex 3D nanostructures.In this review,we emphasize the importance of the criteria for optical performance evaluation of imaging devices,discuss material properties relevant to TPL,fabrication techniques,and highlight the application of TPL in optical imaging.As the first panoramic review on this topic,it will equip researchers with foundational knowledge and recent advancements of TPL for imaging optics,promoting a deeper understanding of the field.By leveraging on its high-resolution capability,extensive material range,and true 3D processing,alongside advances in materials,fabrication,and design,we envisage disruptive solutions to current challenges and a promising incorporation of TPL in future optical imaging applications.
基金supported by the National Natural Science Foundation of China,No.31970906(to WLei)the Natural Science Foundation of Guangdong Province,No.2020A1515011079(to WLei)+4 种基金Key Technologies R&D Program of Guangdong Province,No.2018B030332001(to GC)Science and Technology Projects of Guangzhou,No.202206060002(to GC)the Youth Science Program of the National Natural Science Foundation of China,No.32100793(to ZX)the Pearl River Innovation and Entrepreneurship Team,No.2021ZT09 Y552Yi-Liang Liu Endowment Fund from Jinan University Education Development Foundation。
文摘Over the past decade,a growing number of studies have reported transcription factor-based in situ reprogramming that can directly conve rt endogenous glial cells into functional neurons as an alternative approach for n euro regeneration in the adult mammalian central ne rvous system.Howeve r,many questions remain regarding how a terminally differentiated glial cell can transform into a delicate neuron that forms part of the intricate brain circuitry.In addition,concerns have recently been raised around the absence of astrocyte-to-neuron conversion in astrocytic lineage-tra cing mice.In this study,we employed repetitive two-photon imaging to continuously capture the in situ astrocyte-to-neuron conversion process following ecto pic expression of the neural transcription factor NeuroD1 in both prolife rating reactive astrocytes and lineage-tra ced astrocytes in the mouse cortex.Time-lapse imaging over several wee ks revealed the ste p-by-step transition from a typical astrocyte with numero us short,tapered branches to a typical neuro n with a few long neurites and dynamic growth cones that actively explored the local environment.In addition,these lineage-converting cells were able to migrate ra dially or to ngentially to relocate to suitable positions.Furthermore,two-photon Ca2+imaging and patch-clamp recordings confirmed that the newly generated neuro ns exhibited synchronous calcium signals,repetitive action potentials,and spontaneous synaptic responses,suggesting that they had made functional synaptic connections within local neural circuits.In conclusion,we directly visualized the step-by-step lineage conversion process from astrocytes to functional neurons in vivo and unambiguously demonstrated that adult mammalian brains are highly plastic with respect to their potential for neuro regeneration and neural circuit reconstruction.
基金supported by the National Natural Science Foundation of China(22171001,22305001,51972001,52372073)the Natural Science Foundation of Anhui Province of China(2108085MB49).
文摘The realization of real-time thermal feedback for monitoring photothermal therapy(PTT)under near-infrared(NIR)light irradiation is of great interest and challenge for antitumor therapy.Herein,by assembling highly efficient photothermal conversion gold nanorods and a temperature-responsive probe((E)-4-(4-(diethylamino)styryl)-1-methylpyridin-1-ium,PyS)within MOF-199,an intelligent nanoplatform(AMPP)was fabricated for simultaneous chemodynamic therapy and NIR light-induced temperature-feedback PTT.The fluorescence intensity and temperature of the PyS probe are linearly related due to the restriction of the rotation of the characteristic monomethine bridge.Moreover,the copper ions resulting from the degradation of MOF-199 in an acidic microenvironment can convert H_(2)O_(2)into•OH,resulting in tumor ablation through a Fenton-like reaction,and this process can be accelerated by increasing the temperature.This study establishes a feasible platform for fabricating highly sensitive temperature sensors for efficient temperature-feedback PTT.
基金supported by National Key Research and Development Program of China(No.2022YFA1207600)National Natural Science Foundation of China(Nos.22178395,62175262 and 62005294)。
文摘To overcome the conflict between the long-wavelength excitation and high singlet oxygen quantum yield of photosensitizers,we conjugated a two-photon fluorophore,tetrahydroquinoxaline coumarin(TQ),and an efficient photodynamic therapeutic agent,benzo[a]phenothiazinium(NBS-NH_(2)),through a hexamethylene linker to build a two-photon photosensitizer,TQ-NBS.In TQ-NBS,TQ served as an energy donor and NBS-NH_(2) acted as an energy acceptor;and TQ-NBS was a F?rster resonance energy transfer(FRET)cassette with a 92.8%efficiency.The large two-photon absorption cross-section of TQ allowed photosensitizer TQ-NBS to work in a 900 nm two-photon excitation(TPE)mode,which greatly benefited the deep tissue penetration in PDT treatment.Meanwhile,the excellent phototoxicity and near-infrared fluorescence of NBS-NH2was kept in TQ-NBS under a TPE mode via a FRET process.Photosensitizer TQ-NBS exhibited a high phototoxic efficacy in living cells and tumor-bearing mice.
基金The National Natural Science Foundation of China(No.60678042)the Natural Science Foundation of Jiangsu Province(No.BK2006553)the Pre-Research Project of the National Natural Science Foundation supported by Southeast University(No.9207041399)
文摘Carbazole-core multi-branched chromophores 9-ethyl- 3, 6-bis （ 2- { 4- [ 5- （4-tert-butyl-phenyl） - [ 1, 3, 4 ] oxadiazol-2-yl ] - phenyl }-vinyl） -carbazole（3） and 9-ethyl-3-（ 2- {4-[ 5-（4-tert-butyl- phenyl） -[ 1, 3, 4 ] oxadiazol-2-yl ] -phenyl }-vinyl ） -carbazole （ 2 ） are synthesized through Wittig reaction and characterized by nuclear magnetic resonance（NMR）and infrared（IR）. The two- photon absorption properties of chromophores are investigated. These chromophores exhibit large two-photon absorption crosssections and strong blue two-photon excited fluorescence. The cooperative enhancement of two-photon absorption（TPA） in the multi-branched structures is observed. This enhancement is partly attributed to the electronic coupling between the branches. The electronic push-pull structures in the arm and their cooperative effects help the extended charge transfer for TPA.
基金ACKNOWLEDGMENTS This work was supported by the National Natural Science Foundation of China （No.21073188）.
文摘The wavelength dependence of photoelectron angular distributions （PADs） of two-photon detachment of Cu^- has been directly studied by using the photoelectron map imaging. Results show that for the laser field intensity of 6.0×10^10W/cm^2, PADs exhibit dramatic change with the external field wavelength. Comparison between the experimental observation and the lowest-order perturbation theory prediction indicates that the pattern of PADs can be explained by the interference of the s and d partial waves in the final state. Relative contri- butions of s and d partial waves in the two-photon detachment at different laser wavelengths are obtained.
基金This study was supported by Development Project of Key Laboratory of Big Data Analysis and Knowledge Service of Science and Technology Innovation Platform of Yangzhou-Yangzhou University Cooperation,Grant/Award Number:YBK202204Jiangsu Province 333 High-level Talent.Training Project,Grant/Award Number:BRA2020176。
文摘Backgroud:Thoracic Trauma and Limb Fractures Are the Two most Common Injuries in Multiple Trauma.However,there Is Still a Lack of Mouse Models of Trauma Combining Tibial Shaft Fracture(TSF)and Thoracic Trauma.In this Study,we Attempted to Develop a Novel Mouse Model of TSF Combined with Blunt Chest Trauma(BCT).Methods:A total of 84 C57BL/6J male mice were used as the multiple trauma model.BCT was induced by hitting the chests of mice with heavy objects,and TSF was in-duced by hitting the tibia of mice with heavy objects after intramedullary fixation.Serum specimens of mice were received by cardiac puncture at defined time points of 0,6,12,24,48,and 72 h.Results:Body weight and body temperature tended to decrease within 24 h after mul-tiple trauma.Hemoglobin analyses revealed a decrease during the first 24 h after mul-tiple trauma.Some animals died by cardiac puncture immediately after chest trauma.These animals exhibited the most severe pulmonary contusion and hemorrhage.The level of lung damage varied in diverse mice but was apparent in all animals.Classic he-matoxylin and eosin(H&E)-stained paraffin pulmonary sections of mice with multiple trauma displayed hemorrhage and an immunoinflammatory reaction.Bronchoalveolar lavage fluid(BALF)and serum samples of mice with multiple trauma showed an upreg-ulation of interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-1α(TNF-1α)compared with the control group.Microimaging confirmed the presence of a tibia fracture and pulmonary contusion.Conclusions:The novel mouse multiple trauma model established in this study is a common trauma model that shows similar pathological mechanisms and imaging characteristics in patients with multiple injuries.This study is useful for determining whether blockade or intervention of the cytokine response is beneficial for the treat-ment of patients with multiple trauma.Further research is needed in the future.
文摘Laser processing technologies enable the precise fabrication of arbitrary structures and devices with broad applications in micro-optics,micro-mechanics,and biomedicine.However,its adoption is limited by the large size,complexity,high cost,and low flexibility of optical systems.Metasurfaces enable precise multidimensional control of light fields,aligning well with the development trend toward compact,high-performance optical systems.Here,we review several recent studies on the application of metasurfaces in laser processing technologies,including 3D nanolithography,direct laser writing,and laser cutting.Metasurfaces provide an integrated operational platform with exceptional performance,poised to disrupt conventional laser processing workflows.This combination presents significant cost efficiency and substantial development potential,with promising applications in areas such as imaging,optical storage,advanced sensing,and space on-orbit manufacturing.
基金financial supports from Natural Science Foundation of China(Grant No.61935008,T2325014,62205174 and 62275100)。
文摘Compound eyes(CEs)that feature ultra-compact structures and extraordinary versatility have revealed great potential for cutting-edge applications.However,the optoelectronic integration of CEs with available photodetectors is still challenging because the planar charge-coupled device(CCD)/complementary metal oxide semiconductor(CMOS)detector cannot match the spatially distributed images formed by CE ommatidia.To reach this end,we report here the optoelectronic integration of CEs by manufacturing 3D nonuniform ommatidia for developing an ultra-compact on-chip camera.As a proof-of-concept,we fabricated microscale CEs with uniform and nonuniform ommatidia through femtosecond laser two-photon photopolymerization,and compared their focusing/imaging performance both theoretically and experimentally.By engineering the surface profiles of the ommatidia at different positions of the CE,the images formed by all the ommatidia can be tuned on a plane.In this way,the nonuniform CE can be directly integrated with a commercial CMOS photodetector,forming an ultra-compact CE camera.Additionally,we further combine the CE camera with a microfluidic chip,which can further serve as an on-chip microscopic monitoring system.We anticipate that such an ultra-compact CE camera may find broad applications in microfluidics,robotics,and micro-optics.
基金supported by NIH(R01AG055413),(R01AG085562),(R21AG059134),(R21AG078749),and(S10OD028609)awards(C.R.).NIH Office of the Director,National Institute on Aging.
文摘Alzheimer's disease(AD)is a neurodegenerative disease characterized by a progressive decline in cognitive functions.Given that AD undermines the quality of life for millions and has an extended asymptomatic period,exploring the full AD pathogenesis and seeking the optimal therapeutic solution have become critical and imperative.This allows researchers to intervene,delay,and potentially prevent AD progression.Several clinical imaging methods are utilized routinely to diagnose and monitor AD,such as magnetic resonance imaging(MRI),functional magnetic resonance imaging(fMRI),positron emission tomography(PET),and single photon emission computed tomography(SPECT).Nevertheless,due to their intrinsic drawbacks and restrictions,such as radiation concerns,high cost,long acquisition time,and low spatial resolution,their applications in AD research are limited,especially at the cellular and molecular levels.In contrast,optical microscopic imaging methods overcome these limitations,offering researchers a variety of approaches with distinct advantages to explore AD pathology on diverse models.In this review,we provide a comprehensive overview of commonly utilized optical microscopic imaging techniques in AD research and introduce their contributions to image amyloid beta(Aβ)species.These techniques include fluorescence microscopy(FM),confocal microscopy(CM),two-photon fluorescence microscopy(TPFM),super-resolution microscopy(SRM),expansion microscopy(ExM),and light-sheet fluorescence microscopy(LSFM).In addition,we introduce some related topics,such as the development of near-infrared(NIR)Aβprobes,the Aβplaque hypothesis,and Aβoligomer hypothesis,and the roles of microglia and astrocytes in AD progression.We believe optical microscopic imaging methods continue to play an indispensable role in deciphering the full pathogenesis of AD and advancing therapeutic strategies.
基金supported by the National Key Research and Development Project(Grant No.2023YFB4705300)the National Natural Science Foundation of China(NSFC)(Grant Nos.62205200 and 62375168)the Natural Science Foundation of Shanghai(Grant No.22ZR1431600)。
文摘Highly programmable shape morphing of 4D-printed micro/nanostructures is urgently desired for applications in robotics and intelligent systems.However,due to the lack of autonomous holistic strategies throughout the target shape input,optimal material distribution generation,and fabrication program output,4D nanoprinting that permits arbitrary shape morphing remains a challenging task for manual design.In this study,we report an autonomous inverse encoding strategy to decipher the genetic code for material property distributions that can guide the encoded modeling toward arbitrarily pre-programmed 4D shape morphing.By tuning the laser power of each voxel at the nanoscale,the genetic code can be spatially programmed and controllable shape morphing can be realized through the inverse encoding process.Using this strategy,the 4D-printed structures can be designed and accurately shift to the target morphing of arbitrarily hand-drawn lines under stimulation.Furthermore,as a proof-of-concept,a flexible fiber micromanipulator that can approach the target region through pre-programmed shape morphing is autonomously inversely encoded according to the localized spatial environment.This strategy may contribute to the modeling and arbitrary shape morphing of micro/nanostructures fabricated via 4D nanoprinting,leading to cutting-edge applications in microfluidics,micro-robotics,minimally invasive robotic surgery,and tissue engineering.
文摘Three-dimensional(3D)nanoprinting via two-photon polymerization offers unparalleled design flexibility and precision,thereby enabling rapid prototyping of advanced micro-optical elements and systems that have found important applications in endomicroscopy and biomedical imaging.The potential of this versatile tool for monolithic manufacturing of dynamic micro-opto-electro-mechanical systems(MOEMSs),however,has not yet been sufficiently explored.This work introduces a 3D-nanoprinted lens actuator with a large optical aperture,optimized for remote focusing in miniaturized imaging systems.The device integrates orthoplanar linear motion springs,a self-aligned sintered micro-magnet,and a monolithic lens,actuated by dual microcoils for uniaxial motion.The use of 3D nanoprinting allows complete design freedom for the integrated optical lens,whereas the monolithic fabrication ensures inherent alignment of the lens with the mechanical elements.With a lens diameter of 1.4 mm and a compact footprint of 5.74 mm,it achieves high mechanical robustness at resonant frequencies exceeding 300 Hz while still providing a large displacement range of 200μm(±100μm).A comprehensive analysis of optical and mechanical performance,including the effects of coil temperature and polymer viscoelasticity,demonstrates its advantages over conventional micro-electro-mechanical system actuators,showcasing its potential for next-generation imaging applications.
