查看更多>>摘要：Biological super-resolution microscopy is a new generation of imaging techniques that overcome the～200 nm diffraction limit of conventional light microscopy in spatial resolution.By providing novel spatial or spatiotemporal information on biological processes at nanometer resolution with molecular specificity,it plays an increasingly important role in biomedical sciences.However,its technical con-straints also require trade-offs to balance its spatial resolution,temporal resolution,and light exposure of samples.Recently,deep learning has achieved breakthrough performance in many image processing and computer vision tasks.It has also shown great promise in pushing the performance envelope of biological super-resolution microscopy.In this brief review,we survey recent advances in using deep learning to enhance the performance of biological super-resolution microscopy,focusing primarily on computational reconstruction of super-resolution images.Related key technical challenges are dis-cussed.Despite the challenges,deep learning is expected to play an important role in the development of biological super-resolution microscopy.We conclude with an outlook into the future of this new re-search area.
查看更多>>摘要：Biological super-resolution microscopy is a new generation of imaging techniques that overcome the～200 nm diffraction limit of conventional light microscopy in spatial resolution.By providing novel spatial or spatiotemporal information on biological processes at nanometer resolution with molecular specificity,it plays an increasingly important role in biomedical sciences.However,its technical con-straints also require trade-offs to balance its spatial resolution,temporal resolution,and light exposure of samples.Recently,deep learning has achieved breakthrough performance in many image processing and computer vision tasks.It has also shown great promise in pushing the performance envelope of biological super-resolution microscopy.In this brief review,we survey recent advances in using deep learning to enhance the performance of biological super-resolution microscopy,focusing primarily on computational reconstruction of super-resolution images.Related key technical challenges are dis-cussed.Despite the challenges,deep learning is expected to play an important role in the development of biological super-resolution microscopy.We conclude with an outlook into the future of this new re-search area.
查看更多>>摘要：Fluorescence microscopy has become a routine tool in biology for interrogating life activities with min-imal perturbation.While the resolution of fluorescence microscopy is in theory governed only by the diffraction of light,the resolution obtainable in practice is also constrained by the presence of optical aberrations.The past two decades have witnessed the advent of super-resolution microscopy that overcomes the diffraction barrier,enabling numerous biological investigations at the nanoscale.Adap-tive optics,a technique borrowed from astronomical imaging,has been applied to correct for optical aberrations in essentially every microscopy modality,especially in super-resolution microscopy in the last decade,to restore optimal image quality and resolution.In this review,we briefly introduce the fundamental concepts of adaptive optics and the operating principles of the major super-resolution imaging techniques.We highlight some recent implementations and advances in adaptive optics for active and dynamic aberration correction in super-resolution microscopy.
查看更多>>摘要：Fluorescence microscopy has become a routine tool in biology for interrogating life activities with min-imal perturbation.While the resolution of fluorescence microscopy is in theory governed only by the diffraction of light,the resolution obtainable in practice is also constrained by the presence of optical aberrations.The past two decades have witnessed the advent of super-resolution microscopy that overcomes the diffraction barrier,enabling numerous biological investigations at the nanoscale.Adap-tive optics,a technique borrowed from astronomical imaging,has been applied to correct for optical aberrations in essentially every microscopy modality,especially in super-resolution microscopy in the last decade,to restore optimal image quality and resolution.In this review,we briefly introduce the fundamental concepts of adaptive optics and the operating principles of the major super-resolution imaging techniques.We highlight some recent implementations and advances in adaptive optics for active and dynamic aberration correction in super-resolution microscopy.
查看更多>>摘要：Ferroptosis is a novel form of programmed cell death characterized by iron-dependent lipid peroxida-tion accumulation.It is morphologically,biochemically,and genetically distinct from other known cell death,such as apoptosis,necrosis,and pyroptosis.Its regulatory mechanisms include iron metabolism,fatty acid metabolism,mitochondrial respiration,and antioxidative systems eliminating lipid peroxida-tion,such as glutathione synthesis,selenium-dependent glutathione peroxidase 4,and ubiquinone.The disruption of cellular redox systems causes damage to the cellular membrane leading to ferroptotic cell death.Recent studies have shown that numerous pathological diseases,like tumors,neurodegenera-tive disorders,and ischemia-reperfusion injury are associated with ferroptosis.As such,pharmacologi-cal regulation of ferroptosis either by activation or by suppression will provide a vast potential for treatments of relevant diseases.This review will discuss the advanced progress in ferroptosis and its regulatory mechanisms from both the antioxidative and oxidative sides.In addition,the roles of ferrop-tosis in various tumorigenesis,development,and therapeutic strategies will be addressed,particularly to chemotherapy and immunotherapy,as well as the discoveries from Traditional Chinese Medicine.This review will lead us to have a comprehensive understanding of the future exploration of ferroptos-is and cancer therapy.
查看更多>>摘要：Ferroptosis is a novel form of programmed cell death characterized by iron-dependent lipid peroxida-tion accumulation.It is morphologically,biochemically,and genetically distinct from other known cell death,such as apoptosis,necrosis,and pyroptosis.Its regulatory mechanisms include iron metabolism,fatty acid metabolism,mitochondrial respiration,and antioxidative systems eliminating lipid peroxida-tion,such as glutathione synthesis,selenium-dependent glutathione peroxidase 4,and ubiquinone.The disruption of cellular redox systems causes damage to the cellular membrane leading to ferroptotic cell death.Recent studies have shown that numerous pathological diseases,like tumors,neurodegenera-tive disorders,and ischemia-reperfusion injury are associated with ferroptosis.As such,pharmacologi-cal regulation of ferroptosis either by activation or by suppression will provide a vast potential for treatments of relevant diseases.This review will discuss the advanced progress in ferroptosis and its regulatory mechanisms from both the antioxidative and oxidative sides.In addition,the roles of ferrop-tosis in various tumorigenesis,development,and therapeutic strategies will be addressed,particularly to chemotherapy and immunotherapy,as well as the discoveries from Traditional Chinese Medicine.This review will lead us to have a comprehensive understanding of the future exploration of ferroptos-is and cancer therapy.
查看更多>>摘要：Recently,there has been a lot of interest by using nanobodies (heavy chain-only antibodies produced naturally from the Camelidae) as targeting molecules for molecular imaging,especially for the nuclear medicine imaging.A radiolabeled method that generates a homogeneous product is of utmost impor-tance in radiotracer development for the nuclear medicine imaging.The conventional method for the radiolabeling of nanobodies is non-specifically,which conjugates the radioisotope chelating group to the side chain ε-amine group of lysine or sulfhydryl of cysteine of nanobodies,with a shortcoming of produce of the heterogeneous radiotracer.Here we describe a method for the site-specific radioiso-tope 99mTc labeling of nanobodies by transpeptidase Sortase A.The radiolabeling process includes two steps:first step,NH2-GGGGK(HYNIC)-COOH peptide (GGGGK =NH2-Gly-Gly-Gly-Gly-Lys-COOH,HYNIC =6-hydrazinonicotinyl) was labeled with 99mTc to obtain GGGGK-HYNIC-99mTc;second step,Sortase A catalyzes the formation of a new peptide bond between the peptide motif LPETG (NH2-Leu-Pro-Glu-Thr-Gly-COOH) expressed C-terminally on the nanobody and the N-terminal of GGGGK-HYNIC-99mTc.After a simple purification process,homogeneous single-conjugated and stable 99mTc-labeled nano-bodies were obtained in ＞50％ yield.This approach demonstrates that the Sortase A-mediated conjuga-tion is a valuable strategy for the development of site-specifically 99mTc-labeled nanobodies.
查看更多>>摘要：Recently,there has been a lot of interest by using nanobodies (heavy chain-only antibodies produced naturally from the Camelidae) as targeting molecules for molecular imaging,especially for the nuclear medicine imaging.A radiolabeled method that generates a homogeneous product is of utmost impor-tance in radiotracer development for the nuclear medicine imaging.The conventional method for the radiolabeling of nanobodies is non-specifically,which conjugates the radioisotope chelating group to the side chain ε-amine group of lysine or sulfhydryl of cysteine of nanobodies,with a shortcoming of produce of the heterogeneous radiotracer.Here we describe a method for the site-specific radioiso-tope 99mTc labeling of nanobodies by transpeptidase Sortase A.The radiolabeling process includes two steps:first step,NH2-GGGGK(HYNIC)-COOH peptide (GGGGK =NH2-Gly-Gly-Gly-Gly-Lys-COOH,HYNIC =6-hydrazinonicotinyl) was labeled with 99mTc to obtain GGGGK-HYNIC-99mTc;second step,Sortase A catalyzes the formation of a new peptide bond between the peptide motif LPETG (NH2-Leu-Pro-Glu-Thr-Gly-COOH) expressed C-terminally on the nanobody and the N-terminal of GGGGK-HYNIC-99mTc.After a simple purification process,homogeneous single-conjugated and stable 99mTc-labeled nano-bodies were obtained in ＞50％ yield.This approach demonstrates that the Sortase A-mediated conjuga-tion is a valuable strategy for the development of site-specifically 99mTc-labeled nanobodies.
查看更多>>摘要：Multicolor super-resolution (SR) microscopy plays a critical role in cell biology research and can visual-ize the interactions between different organelles and the cytoskeleton within a single cell.However,more color channels bring about a heavier budget for imaging and sample preparation,and the use of fluorescent dyes of higher emission wavelengths leads to a worse spatial resolution.Recently,deep convolutional neural networks (CNNs) have shown a compelling capability in cell segmentation,super-resolution reconstruction,image restoration,and many other aspects.Taking advantage of CNN's strong representational ability,we devised a deep CNN-based instant multicolor super-resolution im-aging method termed IMC-SR and demonstrated that it could be used to separate different biological components labeled with the same fluorophore,and generate multicolor images from a single super-resolution image in silico.By IMC-SR,we achieved fast three-color live-cell super-resolution imaging with ～100 nm resolution over a long temporal duration,revealing the complicated interactions between multiple organelles and the cytoskeleton in a single COS-7 cell.
查看更多>>摘要：Multicolor super-resolution (SR) microscopy plays a critical role in cell biology research and can visual-ize the interactions between different organelles and the cytoskeleton within a single cell.However,more color channels bring about a heavier budget for imaging and sample preparation,and the use of fluorescent dyes of higher emission wavelengths leads to a worse spatial resolution.Recently,deep convolutional neural networks (CNNs) have shown a compelling capability in cell segmentation,super-resolution reconstruction,image restoration,and many other aspects.Taking advantage of CNN's strong representational ability,we devised a deep CNN-based instant multicolor super-resolution im-aging method termed IMC-SR and demonstrated that it could be used to separate different biological components labeled with the same fluorophore,and generate multicolor images from a single super-resolution image in silico.By IMC-SR,we achieved fast three-color live-cell super-resolution imaging with ～100 nm resolution over a long temporal duration,revealing the complicated interactions between multiple organelles and the cytoskeleton in a single COS-7 cell.
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