查看更多>>摘要：When imaging the nucleus structure of a cell,the out-of-focus fluorescence acts as background and hinders the detection of weak signals.Light-sheet fluorescence microscopy (LSFM) is a wide-field im-aging approach which has the best of both background removal and imaging speed.However,the com-monly adopted orthogonal excitation/detection scheme is hard to be applied to single-cell imaging due to steric hindrance.For LSFMs capable of high spatiotemporal single-cell imaging,the complex instru-ment design and operation largely limit their throughput of data collection.Here,we propose an ap-proach for high-throughput background-free fluorescence imaging of single cells facilitated by the Im-mersion Tilted Light Sheet Microscopy (ImTLSM).ImTLSM is based on a light-sheet projected off the optical axis of a water immersion objective.With the illumination objective and the detection objective placed opposingly,ImTLSM can rapidly patrol and optically section multiple individual cells while maintaining single-molecule detection sensitivity and resolution.Further,the simplicity and robust-ness of ImTLSM in operation and maintenance enables high-throughput image collection to establish background removal datasets for deep learning.Using a deep learning model to train the mapping from epi-illumination images to ImTLSM illumination images,namely PN-ImTLSM,we demonstrated cross-modality fluorescence imaging,transforming the epi-illumination image to approach the background removal performance obtained with ImTLSM.We demonstrated that PN-ImTLSM can be generalized to large-field homogeneous illumination imaging,thereby further improving the imaging throughput.In addition,compared to commonly used background removal methods,PN-ImTLSM showed much better performance for areas where the background intensity changes sharply in space,facilitating high-density single-molecule localization microscopy.In summary,PN-ImTLSM paves the way for background-free fluorescence imaging on ordinary inverted microscopes.
查看更多>>摘要：When imaging the nucleus structure of a cell,the out-of-focus fluorescence acts as background and hinders the detection of weak signals.Light-sheet fluorescence microscopy (LSFM) is a wide-field im-aging approach which has the best of both background removal and imaging speed.However,the com-monly adopted orthogonal excitation/detection scheme is hard to be applied to single-cell imaging due to steric hindrance.For LSFMs capable of high spatiotemporal single-cell imaging,the complex instru-ment design and operation largely limit their throughput of data collection.Here,we propose an ap-proach for high-throughput background-free fluorescence imaging of single cells facilitated by the Im-mersion Tilted Light Sheet Microscopy (ImTLSM).ImTLSM is based on a light-sheet projected off the optical axis of a water immersion objective.With the illumination objective and the detection objective placed opposingly,ImTLSM can rapidly patrol and optically section multiple individual cells while maintaining single-molecule detection sensitivity and resolution.Further,the simplicity and robust-ness of ImTLSM in operation and maintenance enables high-throughput image collection to establish background removal datasets for deep learning.Using a deep learning model to train the mapping from epi-illumination images to ImTLSM illumination images,namely PN-ImTLSM,we demonstrated cross-modality fluorescence imaging,transforming the epi-illumination image to approach the background removal performance obtained with ImTLSM.We demonstrated that PN-ImTLSM can be generalized to large-field homogeneous illumination imaging,thereby further improving the imaging throughput.In addition,compared to commonly used background removal methods,PN-ImTLSM showed much better performance for areas where the background intensity changes sharply in space,facilitating high-density single-molecule localization microscopy.In summary,PN-ImTLSM paves the way for background-free fluorescence imaging on ordinary inverted microscopes.
查看更多>>摘要：Microbiota-host interaction has attracted more and more attentions in recent years.The association between microbiota and host health is largely attributed to its influence on host immune system.Microbial-derived antigens and metabolites play a critical role in shaping the host immune system,including regulating its development,activation,and function.However,during various diseases the microbiota-host communication is frequently found to be disordered.In particular,gut microbiota dysbiosis associated with or led to the occurrence and progression of infectious diseases,autoimmune diseases,metabolic diseases,and neurological diseases.Pathogenic microbes and their metabolites dis-turb the protective function of immune system,and lead to disordered immune responses that usually correlate with disease exacerbation.In the other hand,the immune system also regulates microbiota composition to keep host homeostasis.Here,we will discuss the current advances of our knowledge about the interactions between microbiota and host immune system during health and diseases.
查看更多>>摘要：Microbiota-host interaction has attracted more and more attentions in recent years.The association between microbiota and host health is largely attributed to its influence on host immune system.Microbial-derived antigens and metabolites play a critical role in shaping the host immune system,including regulating its development,activation,and function.However,during various diseases the microbiota-host communication is frequently found to be disordered.In particular,gut microbiota dysbiosis associated with or led to the occurrence and progression of infectious diseases,autoimmune diseases,metabolic diseases,and neurological diseases.Pathogenic microbes and their metabolites dis-turb the protective function of immune system,and lead to disordered immune responses that usually correlate with disease exacerbation.In the other hand,the immune system also regulates microbiota composition to keep host homeostasis.Here,we will discuss the current advances of our knowledge about the interactions between microbiota and host immune system during health and diseases.
查看更多>>摘要：The voltage-dependent cardiac sodium channel plays a key role in cardiac excitability and conduction and it is the drug target of medically important.However,its atomic-resolution structure is still lack.Here,we report a modeled structure of Nav1.5 pore domain in closed state.The structure was con-structed by Rosetta-membrane homology modeling method based on the template of eukaryotic Nav channel NavPaS and selected by energy and direct coupling analysis (DCA).Moreover,this structure was optimized through molecular dynamical simulation in the lipid membrane bilayer.Finally,to vali-date the constructed model,the binding energy and binding sites of closed-state local anesthetics (LAs)in the modeled structure were computed by the MM-GBSA method and the results are in agreement with experiments.The modeled structure of Nav1.5 pore domain in closed state may be useful to ex-plore molecular mechanism of a state-dependent drug binding and helpful for new drug development.
查看更多>>摘要：The voltage-dependent cardiac sodium channel plays a key role in cardiac excitability and conduction and it is the drug target of medically important.However,its atomic-resolution structure is still lack.Here,we report a modeled structure of Nav1.5 pore domain in closed state.The structure was con-structed by Rosetta-membrane homology modeling method based on the template of eukaryotic Nav channel NavPaS and selected by energy and direct coupling analysis (DCA).Moreover,this structure was optimized through molecular dynamical simulation in the lipid membrane bilayer.Finally,to vali-date the constructed model,the binding energy and binding sites of closed-state local anesthetics (LAs)in the modeled structure were computed by the MM-GBSA method and the results are in agreement with experiments.The modeled structure of Nav1.5 pore domain in closed state may be useful to ex-plore molecular mechanism of a state-dependent drug binding and helpful for new drug development.
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