The harm of pathogenic bacteria to humans has promoted extensive research on physiological processes of pathogens,such as the mechanism of bacterial infection,antibiotic mode of action,and bacterial antimicrobial resi...The harm of pathogenic bacteria to humans has promoted extensive research on physiological processes of pathogens,such as the mechanism of bacterial infection,antibiotic mode of action,and bacterial antimicrobial resistance.Most of these processes can be better investigated by timely tracking of fluorophore-derived antibiotics in living cells.In this paper,we will review the recent development of fluorescent antibiotics featuring the conjugation with various fluorophores,and focus on their applications in fluorescent imaging and real-time detection for various physiological processes of bacteria in vivo.展开更多
Backgrounds Time-lapse live cell imaging of a growing cell population is routine in many biological investigations.A major challenge in imaging analysis is accurate segmentation,a process to define the boundaries of c...Backgrounds Time-lapse live cell imaging of a growing cell population is routine in many biological investigations.A major challenge in imaging analysis is accurate segmentation,a process to define the boundaries of cells based on raw image data.Current segmentation methods relying on single boundary features have problems in robustness when dealing with inhomogeneous foci which invariably happens in cell population imaging.Methods:Combined with a multi-layer training set strategy,we developed a neural-network-based algorithm—Cellbow.Results'Cellbow can achieve accurate and robust segmentation of cells in broad and general settings.It can also facilitate long-term tracking of cell growth and division.To facilitate the application of Cellbow,we provide a website on which one can online test the software,as well as an I mage J plugin for the user to visualize the performance before software installation.Conclusions Cellbow is customizable and generalizable.It is broadly applicable to segmenting fluorescent images of diverse cell types with no further training needed.For bright-field images,only a small set of sample images of the specific cell type from the user may be needed for training.展开更多
Safe fluorescent gene-transfection vectors are in great demand for basic biological applications and for gene-therapy research. Here, we introduce a new type of luminescent silicon nanoparticle （SiNP）-based gene car...Safe fluorescent gene-transfection vectors are in great demand for basic biological applications and for gene-therapy research. Here, we introduce a new type of luminescent silicon nanoparticle （SiNP）-based gene carrier suitable for determining the intracellular fate of the gene vehicle in a long-term and real-time manner. The presented SiNP-based nanocarriers simultaneously feature strong and stable fluorescence, high DNA-loading capacity and gene-transfection efficienc35 as well as favorable biocompafibility. Taking advantage of these unique benefits, we were able to readily observe the behavior of the gene carriers in live cells （e.g. cellular uptake, intracellular trafficking, and endosomal escape） in a long-term and real- time manner. The results demonstrate the potential usability of these fluorescent SiNP-based gene vectors as powerful tools in the field of gene therapy, and provide invaluable information for understanding the intracellular behavior of gene carriers.展开更多
基金We are grateful for the financial support from the National Natural Science Foundation of China(21878286,21908216,21576043)Dalian Institute of Chemical Physics(DICPI201938,DICP I202006).
文摘The harm of pathogenic bacteria to humans has promoted extensive research on physiological processes of pathogens,such as the mechanism of bacterial infection,antibiotic mode of action,and bacterial antimicrobial resistance.Most of these processes can be better investigated by timely tracking of fluorophore-derived antibiotics in living cells.In this paper,we will review the recent development of fluorescent antibiotics featuring the conjugation with various fluorophores,and focus on their applications in fluorescent imaging and real-time detection for various physiological processes of bacteria in vivo.
基金This work was supported by the Ministry of Science and Technology of China(2015CB910300)the National Key Research and Development Program of China(2018YFA0900700)the National Natural Science Foundation of China(NSFC31700733).Part of the analysis was performed on the High Performance Computing Platform of the Center for Life Science.
文摘Backgrounds Time-lapse live cell imaging of a growing cell population is routine in many biological investigations.A major challenge in imaging analysis is accurate segmentation,a process to define the boundaries of cells based on raw image data.Current segmentation methods relying on single boundary features have problems in robustness when dealing with inhomogeneous foci which invariably happens in cell population imaging.Methods:Combined with a multi-layer training set strategy,we developed a neural-network-based algorithm—Cellbow.Results'Cellbow can achieve accurate and robust segmentation of cells in broad and general settings.It can also facilitate long-term tracking of cell growth and division.To facilitate the application of Cellbow,we provide a website on which one can online test the software,as well as an I mage J plugin for the user to visualize the performance before software installation.Conclusions Cellbow is customizable and generalizable.It is broadly applicable to segmenting fluorescent images of diverse cell types with no further training needed.For bright-field images,only a small set of sample images of the specific cell type from the user may be needed for training.
基金We express our grateful thanks to Prof. H. J. Dai＇s and Prof. S. T. Lee＇s valuable comments. We also appreciate financial support from the National Basic Research Program of China （Nos. 2013CB934400 and 2012CB932400）, the National Natural Science Foundation of China （Nos. 61361160412, 31400860 and 21575096）, the Natural Science Foundation of Jiangsu Province of China （Nos. BK20130052 and BK20130298）, and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions （PAPD）.
文摘Safe fluorescent gene-transfection vectors are in great demand for basic biological applications and for gene-therapy research. Here, we introduce a new type of luminescent silicon nanoparticle （SiNP）-based gene carrier suitable for determining the intracellular fate of the gene vehicle in a long-term and real-time manner. The presented SiNP-based nanocarriers simultaneously feature strong and stable fluorescence, high DNA-loading capacity and gene-transfection efficienc35 as well as favorable biocompafibility. Taking advantage of these unique benefits, we were able to readily observe the behavior of the gene carriers in live cells （e.g. cellular uptake, intracellular trafficking, and endosomal escape） in a long-term and real- time manner. The results demonstrate the potential usability of these fluorescent SiNP-based gene vectors as powerful tools in the field of gene therapy, and provide invaluable information for understanding the intracellular behavior of gene carriers.
