Wide-field linear structured illumination microscopy(LSIM)extends resolution beyond the diffraction limit by moving unresolvable high-frequency information into the passband of the microscopy in the form of moiré...Wide-field linear structured illumination microscopy(LSIM)extends resolution beyond the diffraction limit by moving unresolvable high-frequency information into the passband of the microscopy in the form of moiréfringes.However,due to the diffraction limit,the spatial frequency of the structured illumination pattern cannot be larger than the microscopy cutoff frequency,which results in a twofold resolution improvement over wide-field microscopes.This Letter presents a novel approach in point-scanning LSIM,aimed at achieving higher-resolution improvement by combining stimulated emission depletion(STED)with point-scanning structured illumination microscopy(ps SIM)(STED-ps SIM).The according structured illumination pattern whose frequency exceeds the microscopy cutoff frequency is produced by scanning the focus of the sinusoidally modulated excitation beam of STED microscopy.The experimental results showed a 1.58-fold resolution improvement over conventional STED microscopy with the same depletion laser power.展开更多
Flexible organic materials that exhibit dynamic ultralong room temperature phosphorescence(DURTP)via photoactivation have attracted increasing research interest for their fascinating functions of reversibly writing-re...Flexible organic materials that exhibit dynamic ultralong room temperature phosphorescence(DURTP)via photoactivation have attracted increasing research interest for their fascinating functions of reversibly writing-reading-erasing graphic information in the form of a long afterglow.However,due to the existence of a nonnegligible activation threshold for the initial exposure dose,the display mode of these materials has thus far been limited to binary patterns.By resorting to halogen element doping of carbon dots(CDs)to enhance intersystem crossing and reduce the activation threshold,we were able to produce,for the first time,a transparent,flexible,and fully programmable DURTP composite film with a reliable grayscale display capacity.Examples of promising applications in UV photography and highly confidential steganography were constructed,partially demonstrating the broad future applications of this material as a programmable platform with a high optical information density.展开更多
Stimulated emission depletion(STED)nanoscopy enables the visualization of subcellular organelles in unprecedented detail.However,reducing the power dependency remains one of the greatest challenges for STED imaging in...Stimulated emission depletion(STED)nanoscopy enables the visualization of subcellular organelles in unprecedented detail.However,reducing the power dependency remains one of the greatest challenges for STED imaging in living cells.Here,we propose a new method,called modulated STED,to reduce the demand for depletion power in STED imaging by modulating the information from the temporal and spatial domains.In this approach,an excitation pulse is followed by a depletion pulse with a longer delay;therefore,the fluorescence decay curve contains both confocal and STED photons in a laser pulse period.With time-resolved detection,we can remove residual diffraction-limited signals pixel by pixel from STED photons by taking the weighted difference of the depleted photons.Finally,fluorescence emission in the periphery of an excitation spot is further inhibited through spatial modulation of fluorescent signals,which replaced the increase of the depletion power in conventional STED.We demonstrate that the modulated STED method can achieve a resolution of<100 nm in both fixed and living cells with a depletion power that is dozens of times lower than that of conventional STED,therefore,it is very suitable for long-term superresolution imaging of living cells.Furthermore,the idea of the method could open up a new avenue to the implementation of other experiments,such as light-sheet imaging,multicolor and three-demensional(3D)super-resolution imaging.展开更多
基金supported by the National Natural Science Foundation of China(Nos.62275168,62275164,61775148,and 61905145)the National Key Research and Development Program of China(No.2022YFA1206300)+5 种基金the Guangdong Natural Science Foundation and Province Project(Nos.2021A1515011916 and 2023A1515012250)the Foundation from Department of Science and Technology of Guangdong Province(No.2021QN02Y124)the Foundation from Department of Education of Guangdong Province(No.2023ZDZX2052)the Shenzhen Science and Technology R&D and Innovation Foundation(No.JCYJ20200109105608771)the Shenzhen Key Laboratory of Photonics and Biophotonics(No.ZDSYS20210623092006020)the Medical-Engineering Interdisciplinary Research Foundation of Shenzhen University。
文摘Wide-field linear structured illumination microscopy(LSIM)extends resolution beyond the diffraction limit by moving unresolvable high-frequency information into the passband of the microscopy in the form of moiréfringes.However,due to the diffraction limit,the spatial frequency of the structured illumination pattern cannot be larger than the microscopy cutoff frequency,which results in a twofold resolution improvement over wide-field microscopes.This Letter presents a novel approach in point-scanning LSIM,aimed at achieving higher-resolution improvement by combining stimulated emission depletion(STED)with point-scanning structured illumination microscopy(ps SIM)(STED-ps SIM).The according structured illumination pattern whose frequency exceeds the microscopy cutoff frequency is produced by scanning the focus of the sinusoidally modulated excitation beam of STED microscopy.The experimental results showed a 1.58-fold resolution improvement over conventional STED microscopy with the same depletion laser power.
基金supported by the National Key R&D Program of China(2021FF0502900)the National Natural Science Foundation of China(61835009,62005171,61975127)+2 种基金the Guangdong Naural Science Foundation(2022A1515011954,2020A1515010679)the Key Project of Guangdong Provincial Deprtment of Education(2021ZDZX2013)the Shenzhen Basic Research Project (JCYJ20180305125304883)。
文摘Flexible organic materials that exhibit dynamic ultralong room temperature phosphorescence(DURTP)via photoactivation have attracted increasing research interest for their fascinating functions of reversibly writing-reading-erasing graphic information in the form of a long afterglow.However,due to the existence of a nonnegligible activation threshold for the initial exposure dose,the display mode of these materials has thus far been limited to binary patterns.By resorting to halogen element doping of carbon dots(CDs)to enhance intersystem crossing and reduce the activation threshold,we were able to produce,for the first time,a transparent,flexible,and fully programmable DURTP composite film with a reliable grayscale display capacity.Examples of promising applications in UV photography and highly confidential steganography were constructed,partially demonstrating the broad future applications of this material as a programmable platform with a high optical information density.
基金This work has been partially supported by the National Basic Research Program of China(No.2017YFA0700500)the National Natural Science Foundation of China(Nos.61620106016,61835009,62005171,and 61975127)+3 种基金Guangdong Natural Science Foundation(Nos.2019A1515110380 and 2020A1515010679)Shenzhen International Cooperation Project(No.GJHZ20180928161811821)Shenzhen Basic Research Project(No.JCYJ20180305125304883)China Post-doctoral Science Foundation(No.2019M663050).
文摘Stimulated emission depletion(STED)nanoscopy enables the visualization of subcellular organelles in unprecedented detail.However,reducing the power dependency remains one of the greatest challenges for STED imaging in living cells.Here,we propose a new method,called modulated STED,to reduce the demand for depletion power in STED imaging by modulating the information from the temporal and spatial domains.In this approach,an excitation pulse is followed by a depletion pulse with a longer delay;therefore,the fluorescence decay curve contains both confocal and STED photons in a laser pulse period.With time-resolved detection,we can remove residual diffraction-limited signals pixel by pixel from STED photons by taking the weighted difference of the depleted photons.Finally,fluorescence emission in the periphery of an excitation spot is further inhibited through spatial modulation of fluorescent signals,which replaced the increase of the depletion power in conventional STED.We demonstrate that the modulated STED method can achieve a resolution of<100 nm in both fixed and living cells with a depletion power that is dozens of times lower than that of conventional STED,therefore,it is very suitable for long-term superresolution imaging of living cells.Furthermore,the idea of the method could open up a new avenue to the implementation of other experiments,such as light-sheet imaging,multicolor and three-demensional(3D)super-resolution imaging.
