Quantitative data analysis in single-molecule localization microscopy(SMLM)is crucial for studying cellular functions at the biomolecular level.In the past decade,several quantitative methods were developed for analyz...Quantitative data analysis in single-molecule localization microscopy(SMLM)is crucial for studying cellular functions at the biomolecular level.In the past decade,several quantitative methods were developed for analyzing SMLM data;however,imaging artifacts in SMLM experiments reduce the accuracy of these methods,and these methods were seldom designed as user-friendly tools.Researchers are now trying to overcome these di±culties by developing easyto-use SMLM data analysis software for certain image analysis tasks.But,this kind of software did not pay su±cient attention to the impact of imaging artifacts on the analysis accuracy,and usually contained only one type of analysis task.Therefore,users are still facing di±culties when they want to have the combined use of different types of analysis methods according to the characteristics of their data and their own needs.In this paper,we report an ImageJ plug-in called DecodeSTORM,which not only has a simple GUI for human–computer interaction,but also combines artifact correction with several quantitative analysis methods.DecodeSTORM includes format conversion,channel registration,artifact correction(drift correction and localization¯ltering),quantitative analysis(segmentation and clustering,spatial distribution statistics and colocalization)and visualization.Importantly,these data analysis methods can be combined freely,thus improving the accuracy of quantitative analysis and allowing users to have an optimal combination of methods.We believe DecodeSTORM is a user-friendly and powerful ImageJ plug-in,which provides an easy and accurate data analysis tool for adventurous biologists who are looking for new imaging tools for studying important questions in cell biology.展开更多
The localization of three monoclonal antibodies (MAb) against gastric cancer was studied on two human gastric cancer cell lines by immunoelectron microscopic technique. It has shown that the corresponding antigens of ...The localization of three monoclonal antibodies (MAb) against gastric cancer was studied on two human gastric cancer cell lines by immunoelectron microscopic technique. It has shown that the corresponding antigens of MAb 3G9 and 3H11 were distributed on the microvilli (M) and non-microvillus (NM) plasma membrane of target cells, with various M to NM ratios depending on the MAbs and target cells used. However, the corresponding antigens of MAb PD4 was only localized on the surface of round or finger-like bulges of target cells and never on the microvilli and non-microvillous plasma membrane. Since the nature and function of these tumor antigens have not been identified yet, the implication of the different distributions of these antigens remians to be clarifated.展开更多
Changes of calmodulin (CaM) distribution in the embryo sac of rice (Oryza sativa subsp. Japonica) at various stages before and after fertilization have been investigated by using immunogold electron microscopy. Before...Changes of calmodulin (CaM) distribution in the embryo sac of rice (Oryza sativa subsp. Japonica) at various stages before and after fertilization have been investigated by using immunogold electron microscopy. Before pollination, both cytoplasm and vacuoles of the egg cell, synergids and central cell were labeled by gold particles. A small amount of gold particles were localized in the nucleus, endoplasmic reticulum, mitochondria and dictyosomes. From pollination to fertilization, CaM amount increased in these cells, especially rich in the starch of amyloplasts. Increase of gold particles in the central cell began about 2 h earlier than that in the egg cell. There was no distinct difference of CaM amount between the degenerated and the persistent synergids. It is interesting to observe an obvious change of CaM distribution form during pollination and fertilization from scattered single particles to clustered particles, and back again to single particles after the fertilization finished. CaM was also localized extracellularly in the embryo sac wall as well as in the wall and intercellular space of nucellus cells. The extracellular CaM also changes in its amount and form after pollination. These results suggest that CaM, either intra- or extra-cellular, may play important roles in fertilization and zygote formation.展开更多
The 3D location and dipole orientation of light emitters provide essential information in many biological,chemical,and physical systems.Simultaneous acquisition of both information types typically requires pupil engin...The 3D location and dipole orientation of light emitters provide essential information in many biological,chemical,and physical systems.Simultaneous acquisition of both information types typically requires pupil engineering for 3D localization and dual-channel polarization splitting for orientation deduction.Here we report a geometric phase helical point spread function for simultaneously estimating the 3D position and dipole orientation of point emitters.It has a compact and simpler optical configuration compared to polarization-splitting techniques and yields achromatic phase modulation in contrast to pupil engineering based on dynamic phase,showing great potential for single-molecule orientation and localization microscopy.展开更多
Traditional technique such nanoindenter（NI） can＇t measure the local elastic modulus at nano-scale（lateral）. Atomic force acoustic microscopy （AFAM） is a dynamic method, which can quantitatively determine indent...Traditional technique such nanoindenter（NI） can＇t measure the local elastic modulus at nano-scale（lateral）. Atomic force acoustic microscopy （AFAM） is a dynamic method, which can quantitatively determine indentation modulus by measuring the contact resonance spectra for high order modes of the cantilever. But there are few reports on the effect of experimental factors, such length of cantilever, contact stiffness on measured value. For three different samples, including copper（Cu） film with 110 nm thickness, zinc（Zn） film of 90 nm thickness and glass slides, are prepared and tested, using referencing approach in which measurements are performed on the test and reference samples （it＇s elastic modulus is known）, and their contact resonance spectra are measured used the AFAM system experimentally. According to the vibration theory, from the lowest two contact resonance frequencies, the tip-sample contact stiffness is calculated, and then the values for the elastic properties of test sample, such as the indentation modulus, are determined. Using AFAM system, the measured indentation modulus of copper nano-film, zinc nano-film and glass slides are 113.53 GPa, 87.92 GPa and 57.04 GPa, which are agreement with literature values Mcu--105-130 GPa, Mzn = 88.44 GPa and Molass = 50-90 GPa. Furthermore, the sensitivity of contact resonance frequency to contact stiffness is analyzed theoretically. The results show that for the cantilevers with the length 160 pm, 225 μm and 520 μm respectively, when contact stiffness increases from 400 N/m to 600 N/m, the increments of first contact resonance frequency are 126 kHz, 93 kHz and 0.6 kHz, which show that the sensitivity of the contact resonance frequency to the contact stiffness reduces with the length of cantilever increasing. The novel method presented can characterize elastic modulus of near surface for nano-film and bulk material, and local elasticity of near surface can be evaluated by optimizing the experimental parameters using the AFAM system.展开更多
Cells employ proteins to perform metabolic functions and maintain active physiological state through charge transfer and energy conversion.These processes are carried out in a narrow space precisely and rapidly,which,...Cells employ proteins to perform metabolic functions and maintain active physiological state through charge transfer and energy conversion.These processes are carried out in a narrow space precisely and rapidly,which,no doubt,bring great difficulty for their detection and dissection.Fortunately,in recent years,the development and expansion of single-molecule technique in protein research make monitoring the dynamical changes of protein at single-molecule level a reality,which also provides a powerful tool for the further exploration of new phenomena and new mechanisms of life activities.This paper aims to summarize the working principle and essential achievements of single-molecule technique in protein research in recent five years.We focus on not only dissecting the difference of nanopores,atomic force microscope,scanning tunneling microscope,and optical tweezers technique,but also discussing the great significance of these single-molecule techniques in investigating intramolecular and intermolecular interactions,electron transport,and conformational changes.Finally,the opportunities and challenges of the single-molecule technique in protein research are discussed,which provide a new door for single-molecule protein research.展开更多
An error correction technique for the micro-scanning instrument of the optical micro-scanning thermal microscope imaging system is proposed. The technique is based on micro-scanning technology combined with the propos...An error correction technique for the micro-scanning instrument of the optical micro-scanning thermal microscope imaging system is proposed. The technique is based on micro-scanning technology combined with the proposed second-order oversampling reconstruction algorithm and local gradient image reconstruction algorithm. In this paper, we describe the local gradient image reconstruction model, the error correction technique, down-sampling model and the error correction principle. In this paper, we use a Lena original image and four low-resolution images obtained from the standard half-pixel displacement to simulate and verify the effectiveness of the proposed technique. In order to verify the effectiveness of the proposed technique, two groups of low-resolution thermal microscope images are collected by the actual thermal microscope imaging system for experimental study. Simulations and experiments show that the proposed technique can reduce the optical micro-scanning errors, improve the imaging effect of the system and improve the system's spatial resolution. It can be applied to other electro-optical imaging systems to improve their resolution.展开更多
BACKGROUND Transduodenal local excision is an alternative treatment approach for benign ampullary tumors.However,this procedure has technical difficulties,especially during reconstruction of the pancreaticobiliary duc...BACKGROUND Transduodenal local excision is an alternative treatment approach for benign ampullary tumors.However,this procedure has technical difficulties,especially during reconstruction of the pancreaticobiliary ducts.An operating microscope has been widely used by surgeons for delicate surgery due to its major advantages of magnification,illumination,and stereoscopic view.The application of an operating microscope in transduodenal excision of ampullary tumors has not been reported.CASE SUMMARY A 55-year-old woman was admitted for investigation of recurrent upper abdominal pain.Physical examination and laboratory tests found no abnormalities.Imaging identified a large mass in the descending part of the duodenum.Esophagogastroduodenoscopy revealed a 3.5-cm-sized villous growth over the major duodenal papilla.Pathology of the endoscopic biopsy indicated a villous adenoma with low-grade dysplasia.Microscopic transduodenal excision of the ampullary tumor was performed.The final pathological diagnosis was villous-tubular adenoma with low-grade dysplasia.The patient was discharged on postoperative day 12 after an uneventful recovery.Endoscopic retrograde cholangiopancreatography was performed 3 mo postoperatively and showed no bile duct or pancreatic duct strictures and no tumor recurrence.The patient is continuing follow-up at our clinic and remains well.CONCLUSION Operating microscope-assisted transduodenal local excision is a feasible and effective option for benign ampullary tumors.展开更多
为了研究砂岩在渗流−应力耦合作用下的变形局部化破坏特征,利用可视化三轴伺服控制试验系统,结合三维数字图像相关技术(three-dimensional digital image correlation,简称3D-DIC),开展不同排水条件下的砂岩三轴压缩试验,对岩石力学、...为了研究砂岩在渗流−应力耦合作用下的变形局部化破坏特征,利用可视化三轴伺服控制试验系统,结合三维数字图像相关技术(three-dimensional digital image correlation,简称3D-DIC),开展不同排水条件下的砂岩三轴压缩试验,对岩石力学、渗流及其变形局部化特性进行分析,并利用电镜扫描对砂岩破坏后其破裂面微观形貌进行分析。结果表明:排水条件下砂岩的峰值强度和弹性模量高于不排水条件下的值,且随着渗透水压增大,砂岩的峰值强度、弹性模量和泊松比均随之增大,出现贯通裂纹的时间点和渗透率最大值出现的时间点则会提前;当渗透水压相同时,不排水条件下砂岩表面变形场云图的局部化带比排水条件下的更宽,即岩石的宏观裂纹更明显,排水条件下水流将岩石内部的矿物颗粒带走,形成孔洞,其破裂面的表面比不排水条件下更光滑,而不排水条件下颗粒表面明显有片状岩屑附着;所有排水条件下径向变形局部化启动点始终高于轴向变形局部化启动点,平均提高了1.23%,变形局部化径向、轴向启动应力水平均随渗透水压的增大而增大,即启动的时间点更提前,排水条件下砂岩的径向启动应力水平、轴向启动应力水平均高于不排水条件下的值,平均分别提高了1.85%和2.21%,渗透水压相同时,不排水条件下启动应力及应力水平受水压的影响比排水条件更明显。展开更多
基金supported by the National Natural Science Foundation of China(82160345)Key research and development project of Hainan province(ZDYF2021GXJS017)+2 种基金Key Science and Technology Plan Project of Haikou(2021-016)the Start-up Fund from Hainan University(KYQD(ZR)-20022 and KYQD(ZR)-20077)the Student Innovation and Entrepreneurship Project of Biomedical Engineer-ing School,Hainan University(BMECF2D2021001).
文摘Quantitative data analysis in single-molecule localization microscopy(SMLM)is crucial for studying cellular functions at the biomolecular level.In the past decade,several quantitative methods were developed for analyzing SMLM data;however,imaging artifacts in SMLM experiments reduce the accuracy of these methods,and these methods were seldom designed as user-friendly tools.Researchers are now trying to overcome these di±culties by developing easyto-use SMLM data analysis software for certain image analysis tasks.But,this kind of software did not pay su±cient attention to the impact of imaging artifacts on the analysis accuracy,and usually contained only one type of analysis task.Therefore,users are still facing di±culties when they want to have the combined use of different types of analysis methods according to the characteristics of their data and their own needs.In this paper,we report an ImageJ plug-in called DecodeSTORM,which not only has a simple GUI for human–computer interaction,but also combines artifact correction with several quantitative analysis methods.DecodeSTORM includes format conversion,channel registration,artifact correction(drift correction and localization¯ltering),quantitative analysis(segmentation and clustering,spatial distribution statistics and colocalization)and visualization.Importantly,these data analysis methods can be combined freely,thus improving the accuracy of quantitative analysis and allowing users to have an optimal combination of methods.We believe DecodeSTORM is a user-friendly and powerful ImageJ plug-in,which provides an easy and accurate data analysis tool for adventurous biologists who are looking for new imaging tools for studying important questions in cell biology.
文摘The localization of three monoclonal antibodies (MAb) against gastric cancer was studied on two human gastric cancer cell lines by immunoelectron microscopic technique. It has shown that the corresponding antigens of MAb 3G9 and 3H11 were distributed on the microvilli (M) and non-microvillus (NM) plasma membrane of target cells, with various M to NM ratios depending on the MAbs and target cells used. However, the corresponding antigens of MAb PD4 was only localized on the surface of round or finger-like bulges of target cells and never on the microvilli and non-microvillous plasma membrane. Since the nature and function of these tumor antigens have not been identified yet, the implication of the different distributions of these antigens remians to be clarifated.
文摘Changes of calmodulin (CaM) distribution in the embryo sac of rice (Oryza sativa subsp. Japonica) at various stages before and after fertilization have been investigated by using immunogold electron microscopy. Before pollination, both cytoplasm and vacuoles of the egg cell, synergids and central cell were labeled by gold particles. A small amount of gold particles were localized in the nucleus, endoplasmic reticulum, mitochondria and dictyosomes. From pollination to fertilization, CaM amount increased in these cells, especially rich in the starch of amyloplasts. Increase of gold particles in the central cell began about 2 h earlier than that in the egg cell. There was no distinct difference of CaM amount between the degenerated and the persistent synergids. It is interesting to observe an obvious change of CaM distribution form during pollination and fertilization from scattered single particles to clustered particles, and back again to single particles after the fertilization finished. CaM was also localized extracellularly in the embryo sac wall as well as in the wall and intercellular space of nucellus cells. The extracellular CaM also changes in its amount and form after pollination. These results suggest that CaM, either intra- or extra-cellular, may play important roles in fertilization and zygote formation.
基金supported by the National Natural Science Foundation of China(Nos.62105368,62275268,and 62375284)the Science and Technology Innovation Program of Hunan Province(No.2023RC3010)。
文摘The 3D location and dipole orientation of light emitters provide essential information in many biological,chemical,and physical systems.Simultaneous acquisition of both information types typically requires pupil engineering for 3D localization and dual-channel polarization splitting for orientation deduction.Here we report a geometric phase helical point spread function for simultaneously estimating the 3D position and dipole orientation of point emitters.It has a compact and simpler optical configuration compared to polarization-splitting techniques and yields achromatic phase modulation in contrast to pupil engineering based on dynamic phase,showing great potential for single-molecule orientation and localization microscopy.
基金supported by National Natural Science Foundation of China(Grant No. 50775005)General Program of Science and Technology Development Project of Beijing Municipal Education Commission(Grant No. KM201110015009)
文摘Traditional technique such nanoindenter（NI） can＇t measure the local elastic modulus at nano-scale（lateral）. Atomic force acoustic microscopy （AFAM） is a dynamic method, which can quantitatively determine indentation modulus by measuring the contact resonance spectra for high order modes of the cantilever. But there are few reports on the effect of experimental factors, such length of cantilever, contact stiffness on measured value. For three different samples, including copper（Cu） film with 110 nm thickness, zinc（Zn） film of 90 nm thickness and glass slides, are prepared and tested, using referencing approach in which measurements are performed on the test and reference samples （it＇s elastic modulus is known）, and their contact resonance spectra are measured used the AFAM system experimentally. According to the vibration theory, from the lowest two contact resonance frequencies, the tip-sample contact stiffness is calculated, and then the values for the elastic properties of test sample, such as the indentation modulus, are determined. Using AFAM system, the measured indentation modulus of copper nano-film, zinc nano-film and glass slides are 113.53 GPa, 87.92 GPa and 57.04 GPa, which are agreement with literature values Mcu--105-130 GPa, Mzn = 88.44 GPa and Molass = 50-90 GPa. Furthermore, the sensitivity of contact resonance frequency to contact stiffness is analyzed theoretically. The results show that for the cantilevers with the length 160 pm, 225 μm and 520 μm respectively, when contact stiffness increases from 400 N/m to 600 N/m, the increments of first contact resonance frequency are 126 kHz, 93 kHz and 0.6 kHz, which show that the sensitivity of the contact resonance frequency to the contact stiffness reduces with the length of cantilever increasing. The novel method presented can characterize elastic modulus of near surface for nano-film and bulk material, and local elasticity of near surface can be evaluated by optimizing the experimental parameters using the AFAM system.
基金This work was financially supported by the National Natural Science Foundation of China(No.21978245)National Postdoctoral Program for Innovative Talents(No.BX20200197).
文摘Cells employ proteins to perform metabolic functions and maintain active physiological state through charge transfer and energy conversion.These processes are carried out in a narrow space precisely and rapidly,which,no doubt,bring great difficulty for their detection and dissection.Fortunately,in recent years,the development and expansion of single-molecule technique in protein research make monitoring the dynamical changes of protein at single-molecule level a reality,which also provides a powerful tool for the further exploration of new phenomena and new mechanisms of life activities.This paper aims to summarize the working principle and essential achievements of single-molecule technique in protein research in recent five years.We focus on not only dissecting the difference of nanopores,atomic force microscope,scanning tunneling microscope,and optical tweezers technique,but also discussing the great significance of these single-molecule techniques in investigating intramolecular and intermolecular interactions,electron transport,and conformational changes.Finally,the opportunities and challenges of the single-molecule technique in protein research are discussed,which provide a new door for single-molecule protein research.
基金Supported by Postgraduate Innovation Funding Project of Hebei Province(CXZZSS2019050)the Qinhuangdao City Key Research and Development Program Science and Technology Support Project(201801B010)
文摘An error correction technique for the micro-scanning instrument of the optical micro-scanning thermal microscope imaging system is proposed. The technique is based on micro-scanning technology combined with the proposed second-order oversampling reconstruction algorithm and local gradient image reconstruction algorithm. In this paper, we describe the local gradient image reconstruction model, the error correction technique, down-sampling model and the error correction principle. In this paper, we use a Lena original image and four low-resolution images obtained from the standard half-pixel displacement to simulate and verify the effectiveness of the proposed technique. In order to verify the effectiveness of the proposed technique, two groups of low-resolution thermal microscope images are collected by the actual thermal microscope imaging system for experimental study. Simulations and experiments show that the proposed technique can reduce the optical micro-scanning errors, improve the imaging effect of the system and improve the system's spatial resolution. It can be applied to other electro-optical imaging systems to improve their resolution.
基金Supported by Natural Science Foundation of Zhejiang Province,No.LQ19H100004.
文摘BACKGROUND Transduodenal local excision is an alternative treatment approach for benign ampullary tumors.However,this procedure has technical difficulties,especially during reconstruction of the pancreaticobiliary ducts.An operating microscope has been widely used by surgeons for delicate surgery due to its major advantages of magnification,illumination,and stereoscopic view.The application of an operating microscope in transduodenal excision of ampullary tumors has not been reported.CASE SUMMARY A 55-year-old woman was admitted for investigation of recurrent upper abdominal pain.Physical examination and laboratory tests found no abnormalities.Imaging identified a large mass in the descending part of the duodenum.Esophagogastroduodenoscopy revealed a 3.5-cm-sized villous growth over the major duodenal papilla.Pathology of the endoscopic biopsy indicated a villous adenoma with low-grade dysplasia.Microscopic transduodenal excision of the ampullary tumor was performed.The final pathological diagnosis was villous-tubular adenoma with low-grade dysplasia.The patient was discharged on postoperative day 12 after an uneventful recovery.Endoscopic retrograde cholangiopancreatography was performed 3 mo postoperatively and showed no bile duct or pancreatic duct strictures and no tumor recurrence.The patient is continuing follow-up at our clinic and remains well.CONCLUSION Operating microscope-assisted transduodenal local excision is a feasible and effective option for benign ampullary tumors.
文摘为了研究砂岩在渗流−应力耦合作用下的变形局部化破坏特征,利用可视化三轴伺服控制试验系统,结合三维数字图像相关技术(three-dimensional digital image correlation,简称3D-DIC),开展不同排水条件下的砂岩三轴压缩试验,对岩石力学、渗流及其变形局部化特性进行分析,并利用电镜扫描对砂岩破坏后其破裂面微观形貌进行分析。结果表明:排水条件下砂岩的峰值强度和弹性模量高于不排水条件下的值,且随着渗透水压增大,砂岩的峰值强度、弹性模量和泊松比均随之增大,出现贯通裂纹的时间点和渗透率最大值出现的时间点则会提前;当渗透水压相同时,不排水条件下砂岩表面变形场云图的局部化带比排水条件下的更宽,即岩石的宏观裂纹更明显,排水条件下水流将岩石内部的矿物颗粒带走,形成孔洞,其破裂面的表面比不排水条件下更光滑,而不排水条件下颗粒表面明显有片状岩屑附着;所有排水条件下径向变形局部化启动点始终高于轴向变形局部化启动点,平均提高了1.23%,变形局部化径向、轴向启动应力水平均随渗透水压的增大而增大,即启动的时间点更提前,排水条件下砂岩的径向启动应力水平、轴向启动应力水平均高于不排水条件下的值,平均分别提高了1.85%和2.21%,渗透水压相同时,不排水条件下启动应力及应力水平受水压的影响比排水条件更明显。
